AU740663B2 - Aminoalkyl glucosamine phosphate compounds and their use as adjuvants and immunoeffectors - Google Patents

Description

WO 98/50399 PCT/US98/09385 1

DESCRIPTION

AMINOALKYL GLUCOSAMINE PHOSPHATE COMPOUNDS AND THEIR USE AS ADJUVANTS AND IMMUNOEFFECTORS Background of the Invention Humoral immunity and cell-mediated immunity are the two major branches of the mammalian immune response. Humoral immunity involves the generation of antibodies to foreign antigens. Antibodies are produced by B-lymphocytes. Cell-mediated immunity involves the activation of T-lymphocytes which either act upon infected cells bearing foreign antigens or stimulate other cells to act upon infected cells. Both branches of the mammalian immune system are important in fighting disease. Humoral immunity is the major line of defense against bacterial pathogens. In the case of viral disease, the induction of cytotoxic T lymphocytes (CTLs) appears to be crucial for protective immunity. An effective vaccine stimulates both branches of the immune system to protect against disease.

Vaccines present foreign antigens from disease causing agents to a host so that the host can mount a protective immune response. Often vaccine antigens are killed or attenuated forms of the microbes which cause the disease. The presence of non-essential components and antigens in these killed or attenuated vaccines has encouraged considerable efforts to refine vaccine components including developing well-defined synthetic antigens using chemical and recombinant techniques. The refinement and simplification of microbial vaccines, however, has led to a concomitant loss in potency.

Low-molecular weight synthetic antigens, though devoid of potentially harmful contaminants, are themselves not very immunogenic. These observations have led investigators to add adjuvants to vaccine compositions to potentiate the activity of the refined vaccine components.

Presently, the only adjuvant licensed for human use in the United States is alum, a group of aluminum salts aluminum hydroxide, aluminum phosphate) in which vaccine antigens are formulated. Particulate carriers like alum serve to promote the WO 98/50399 PCT/US98/09385 2 uptake, processing and presentation of soluble antigens by macrophage. Alum, however, is not without side-effects and enhances humoral (antibody) immunity only.

An effective adjuvant potentiates both a humoral and cellular immune response in vaccinated animals. Further, an adjuvant must enhance a host's natural immune response and not aggravate the host system. A well-defined synthetic adjuvant free from extraneous matter which is stable and easy to manufacture would provide these qualities.

Compounds that have been prepared and tested for adjuvanticity (Shimizu et al. 1985, Bulusu et al. 1992, Ikeda et al. 1993, Shimizu et al. 1994, Shimizu et al. 1995, Miyajima et al. 1996), however, often display toxic properties, are unstable and/or have unsubstantial immunostimulatory effects.

The discovery and development of effective adjuvants is essential for improving the efficacy and safety of existing vaccines. Adjuvants impart synthetic peptide and carbohydrate antigens with sufficient immunogenicity to insure the success of the synthetic vaccine approach. There remains a need for new compounds having potent immunomodulating effects.

Summary of the Invention The compounds of the subject invention are aminoalkyl glucosamine phosphate compounds (AGPs) which are adjuvants and immunoeffectors. An aminoalkyl (aglycon) group is glycosidically linked to a 2 -deoxy-2-amino-a-D-glucopyranose (glucosamine) to form the basic structure of the claimed molecules. The compounds are phosphorylated at the 4 or 6 carbon on the glucosamine ring. Further, the compounds possess three 3alkanoyloxyalkanoyl residues.

The compounds of the subject invention are immunoeffector molecules augmenting antibody production in immunized animals, stimulating cytokine production and activating macrophage. In accordance with the subject invention, methods for using these compounds as adjuvants and immunoeffectors are disclosed.

Detailed Description of the Invention The compounds of the subject invention are adjuvant and immunoeffector molecules which are aminoalkyl glucosamine phosphates (AGPs). The compounds WO 98/50399 PCT/US98/09385 3 comprise a 2 -deoxy-2-amino-a-D-glucopyranose (glucosamine) in glycosidic linkage with an aminoalkyl (aglycon) group. Compounds are phosphorylated at the 4 or 6 carbon on the glucosamine ring and have three alkanoyloxyalkanoyl residues. The compounds of the subject invention are described generally by Formula I,

OR,

R0 (CNH

R

4 (CH) 0

C

4 (C4) (Cl4)

(I)

wherein X represents an oxygen or sulfur atom, Y represents an oxygen atom or NH group, and are integers from 0 to 6, R 2 and R 3 represent normal fatty acyl residues having 7 to 16 carbon atoms, R 4 and R 5 are hydrogen or methyl, R 6 and

R

7 are hydrogen, hydroxy, alkoxy, phosphono, phosphonooxy, sulfo, sulfooxy, amino, mercapto, cyano, nitro, formyl or carboxy and esters and amides thereof; R, and R, are phosphono or hydrogen. The configuration of the 3' stereogenic centers to which the normal fatty acyl residues are attached is R or S, but preferably R. The stereochemistry of the carbon atoms to which R 4 or R 5 are attached can be R or S. All stereoisomers, both enantiomers and diastereomers, and mixtures thereof, are considered to fall within the scope of the subject invention.

The heteroatom X of the compounds of the subject invention can be oxygen or sulfur. In a preferred embodiment, X is oxygen. Although the stability of the molecules WO 98/50399 PCT/US98/09385 4 could be effected by a substitution at X, the immunomodulating activity of molecules with these substitutions is not expected to change.

The number of carbon atoms between heteroatom X and the aglycon nitrogen atom is determined by variables and Variables and can be integers from 0 to 6. In a preferred embodiment, the total number of carbon atoms between heteroatom X and the aglycon nitrogen atom is from about 2 to about 6 and most preferably from about 2 to about 4.

The compounds of the subject invention are aminoalkyl glucosamine compounds which are phosphorylated. Compounds can be phosphorylated at position 4 or 6 (R 8 or

R

9 on the glucosamine ring and are most effective if phosphorylated on at least one of these positions. In a preferred embodiment, R 8 is phosphono and R9 is hydrogen.

The compounds of the subject invention are hexaacylated, that is they contain a total of six fatty acid residues. The aminoalkyl glucosamine moiety is acylated at the 2amino and 3-hydroxyl groups of the glucosamine unit and at the amino group of the aglycon unit with 3-hydroxyalkanoyl residues. In Formula I, these three positions are acylated with 3-hydroxytetradecanoyl moieties. The 3-hydroxytetradecanoyl residues are, in turn, substituted with normal fatty acids (Ri-R 3 providing three 3-nalkanoyloxytetradecanoyl residues or six fatty acid groups in total.

The chain length of normal fatty acids Ri-R 3 can be from about 7 to about 16 carbons. Preferably, R,-R 3 are from about 9 to about 14 carbons. The chain lengths of these normal fatty acids can be the same or different. Although, only normal fatty acids are described, it is expected that unsaturated fatty acids fatty acid moieties having double or triple bonds) substituted at R 1

-R

3 on the compounds of the subject invention would produce biologically active molecules. Further, slight modifications in the chain length of the 3-hydroxyalkanoyl residues are not expected to dramatically effect biological activity.

The compounds of the subject invention are adjuvants and immunoeffectors which enhance the generation of antibody in immunized animals, stimulate the production of cytokines and stimulate a cell-mediated immune response including a cytotoxic T-lymphocyte response. In methods for effecting the immune response of an individual, the compounds of the subject invention can be formulated with a WO 98/50399 PCTIUS98/09385 pharmaceutically acceptable carrier for injection or ingestion. As used herein, "pharmaceutically acceptable carrier" means a medium which does not interfere with the immunomodulatory activity of the active ingredient and is not toxic to the patient to whom it is administered. Pharmaceutically acceptable carriers include oil-in-water or water-in-oil emulsions, aqueous compositions, liposomes, microbeads and microsomes.

Formulations of the compounds of the subject invention that can be administered parenterally, i.e. intraperitoneally, subcutaneously or intramuscularly include the following preferred carriers. Examples of preferred carriers for subcutaneous use include a phosphate buffered saline (PBS) solution and 0.01-0.1 triethanolamine in USP Water for Injection. Suitable carriers for intramuscular injection include 10% USP ethanol, propylene glycol and the balance an acceptable isotonic solution such as dextrose. Examples of preferred carriers for intravenous use include 10% USP ethanol, 40% USP propylene glycol and the balance USP Water for Injection. Another acceptable carrier includes 10% USP ethanol and USP Water for Injection; yet another acceptable carrier is 0.01-0.1% triethanolamine in USP Water for Injection.

Pharmaceutically acceptable parenteral solvents are such as to provide a solution or dispersion may be filtered through a 5 micron filter without removing the active ingredient.

Examples of carriers for administration via mucosal surfaces depend upon the particular route. When administered orally, pharmaceutical grades of mannitol, starch, lactose, magnesium stearate, sodium saccharide, cellulose, magnesium carbonate and the like, with mannitol being preferred. When administered intranasally, polyethylene glycol or glycols, sucrose, and/or methylcellulose, and preservatives such as benzalkonium chloride, EDTA, may be used, with polyethylene glycols being preferred, and when administered by inhalation, suitable carriers are polyethylene glycol or glycols, methylcellulose, dispensing agents, and preservatives, with polyethylene glycols being preferred.

The compounds of the subject invention are administered to an individual in "an effective amount" to effect or enhance the individual's immune response. As used herein, "an effective amount" is that amount which shows a response over and above the vehicle WO 98/50399 PCT/US98/09385 6 or negative controls. The precise dosage of the compounds of the subject invention to be administered to a patient will depend upon the particular AGP used, the route of administration, the pharmaceutical composition, and the patient. For example, when administered subcutaneously to enhance an antibody response, the amount of AGP used is from 1 to about 250 micrograms, preferably from about 25 to about 50 micrograms based upon administration to a typical 70 kg adult patient.

In vaccine compositions, the AGPs of the subject invention are administered to a warm-blooded animal, including humans, with an antigen. The amount of antigen administered to elicit a desired response can be readily determined by one skilled in the art and will vary with the type of antigen administered, route of administration and immunization schedule. For example, 0.2 ug of tetanus toxoid administered with the claimed compounds subcutaneously to a mouse in two immunization 21 days apart elicited a humoral immune response to that antigen.

The compounds of the subject invention are synthesized by coupling an Nacyloxyacylated or N-protected aminoalkanol or aminoalkanethiol (aglycon unit) with a suitably protected and/or 3-O-acyloxyacylated glucosamine unit. In one preferred method for preparing the compounds of the subject invention (Scheme an N-(2,2,2trichloroethoxycarbonyl (Troc))-protected glycosyl halide 1 (Z F, Cl, Br) is coupled with an N-[(R)-3-n-alkanoyloxytetradecanoyl]aminoalkanol or thiol 2 (possessing R and

R

7 in suitably protected form) via a Koenigs-Knorr type reaction in the presence of mercury or silver salts to give glycoside intermediate 3. Preferably, the glucosamine unit 1 possesses an anomeric chloride atom (Z Cl), and the coupling catalyst is silver trifluoromethanesulfonate. Intermediate 3 can also be prepared by coupling the aglycon unit 2 with an N-Troc-protected glycosyl acetate (Z OAc) or related activated derivative in the presence of a Lewis acid such as boron trifluoride etherate. By "activated" is meant having an appropriate displaceable leaving group attached to the anomeric center of the glucosamine unit. Glucosamine unit 1 bears an alkanoyloxytetradecanoyl residue on the 3-position, and suitable protecting groups on the 6-hydroxyl and 4-phosphate moieties. Typical protecting groups for the phosphate group include, but are not limited to, phenyl, benzyl, and o-xylyl. The phosphate group is protected preferably with two phenyl groups. The 6-position can be temporarily WO 98/50399 PCT/US98/09385 7 protected by blocking groups commonly used in sugar chemistry such as silyl, benzyl, or benzyloxymethyl ethers or, alternatively, an alkyl carbonate. The 6-hydroxyl group is protected preferably as a 1,1 -dimethyl-2,2,2-trichloroethyl carbonate (TCBOC).

The trichloroethyl-based protecting group(s) in the Koenigs-Knorr coupled product 3 are removed with zinc and the glucosamine nitrogen is selectively acylated with a (R)-3-n-alkanoyloxytetradecanoic acid 4 in the presence of a suitable coupling reagent to give the hexaacylated derivative 5. The remaining protecting groups in 5 are then cleaved by catalytic hydrogenation in the presence of a palladium or platinum catalyst or by other appropriate means to give compounds of Formula A suitable starting material for the synthesis of glycosyl donor 1 is 2- (trimethylsilyl)ethyl 2 -amino- 2 -deoxy-4,6-O-isopropylidene-p-D-glucopyranoside which can be prepared from commercially available D-glucosamine hydrochloride using published procedures. The conversion of the 2-(trimethylsilyl)ethyl glycoside starting material to glycosyl donor 1 can be achieved by methods known in the art or modifications thereof which are described herein. The aglycon unit 2 can be prepared by N-acyloxyacylation of commercially available starting materials with an appropriate (R)-3-n-alkanoyloxytetradecanoic acid 4, or N-acyloxyacylation of starting materials that can be obtained by known methods in the chemical literature. Alternatively, the Nacyloxyacyl residue in 2 can be substituted with an appropriate amine protecting group which is removed subsequent to the coupling reaction such as is described in the second preferred embodiment below.

In a second preferred method for preparing the compounds of the subject invention (Scheme introduction of the (R)-3-n-alkanoyloxytetradecanoyl and phosphate groups into the glucosamine and aglycon units is performed subsequent to the glycosylation (coupling) reaction using N- and O-protecting groups suitable for the chemical differentiation of the amino and hydroxyl groups present. Preferably, the N- Troc-protected glycosyl donor 6 is coupled with anN-allyloxycarbonyl (AOC)-protected aminoalkanol or thiol 7 in the presence of an appropriate catalyst to give the aminoalkyl P-glycoside 8. Most preferably, the glycosyl donor 6 possesses an anomeric acetoxy group (Z OAc), and the coupling catalyst is boron trifluoride etherate. Other Nprotecting groups for the aglycon amino group include, but are not limited to, WO 98/50399 PCT/US98/09385 8 commonly employed carbamates obvious to one skilled in the art such as t-butyl (t- BOC), benzyl (Cbz), 2,2,2-trichloroethyl (Troc), and 9-fluorenylmethyl(Fmoc).

Base-induced cleavage of the acetate groups in coupling product 8 and 4,6acetonide formation under standard conditions known in the art gives intermediate 9.

3-O-Acylation of 9 with (R)-3-n-alkanoyloxytetradecanoic acid 4, followed by palladium(0)-mediated removal of the aglycon N-AOC group and N-acylation with 3-n-alkanoyloxytetradecanoic acid 4 provides intermediate 10. Acetonide hydrolysis and functionalization of the 4- and 6-positions as described herein for the preparation of glycosyl donor 1 gives intermediate 3 O) which is then processed as in Scheme 1 to afford compounds of general Formula The present invention is further described by way of the following non-limiting Examples and Test Examples which are given for illustrative purposes only. It is important to note that the introduction of the (R)-3-n-alkanoyloxytetradecanoyl groups and the phosphate group(s) into the glucosamine and aglycon units do not necessarily have to be performed in the order shown in Schemes 1 and 2 or described in the Examples shown below.

WO 98/50399 WO 9850399PCTIUS98/09385 (PhO) 2 P- 0 Troc n-CI IH23 I Z halide, OAc, etc,

NH

EX 4~4~ N q R 7

R

4 (clV 2 )p 220

H

6 -4OR 3 n-C 1 1 H2 catalyst 100 (PhO) 2 P-O 0R NH R 4

(CH

2 )p RIOm Troc 1 OR n-CIIH23 X6 n-C 1

O

OH

0 I 1 (PhO) 2

P-O

O NH

R

4 (CI 2 ~0R n-C 1 I2 n-C 1 H23 1. Zn, AcOH 2. 4 R=R 2

OR

fl-C,,IH"'

CO

2

H

4 R=R 1

,R

2 orR 3 Scheme 1 WO 98/50399 WO 9850399PCTIUS98/09385 HX I-q

R

7 R AO AcO.

catalyst 6 Z halide, QAc, etc.

QAc ACO 0oR 1. NH 4 OH,. meOH NH Y q R 7 27 Me2C(OMe),kH Trac

I

R

6 0 0 0 R5 1 4 R=R 1 HO qR 2.Pd(O) NHc R AOC 3.4 R=R 3 9 aq. AcOH NHl R4(H e= 2. TCBOC-Cl Troc 3. (PhO)2P(O)CI n- 1 1 ~10 n-C IH23

OR

n-C, H3 1 C2 4 R =RI, R 2 or R 3 Scheme 2 WO 98/50399 PCT/US98/09385 11 Examples 1-29 describe methods of making the AGP compounds of the subject invention. Test Examples 1-7 describe assays conducted to the determine the immunogenicity of these compounds. Table 1 lists the chemical composition and experimental reference numbers for each compound in these examples.

Table 1.

Example Ref. No. R 1

R

3 n p R 6 q R 7 0 2 B1* n-C 13

H

27 C0 0 1 OH 0 H 3 B2** n-C 1 3 H 27 CO 0 1 OH 0 H 4 B3 n-C,,H 23 C0 0 1 OH 0 H B4 i7-C 10

H,

1 C0 0 1 OH 0 H 6 B5 n-C 9

H

19 C0 0 1 OH 0 H 7 n-C 9

H

19 C0 0 1 OH 0 H 8 B7 n-C 8

H

17 C0 0 .1 OH 0 H 9 B8 n-C 6

H

13 C0 0 1 OH 0 H B9 n-C 9

H

1 9 C0 1 1 OH 0 H I1I BIO n-C 9

H

19 C0 0 2 OH 0 H 12 B11 n-C 13

H

27 C0 0 0 CO 2 H 0 H 13B12 n-C, 1

H

23 C0 0 0 CO 2 H 0 H 14 B13 n-C 10

H,

1 C0 0 0 CO 2 H 0 H B14** n-C 9

H

1 9 C0 0 0 CO.,H 0 H0 16 B15* n-C 9 Hl 9 C0 0 0 CO2H 0 H Table I continued.

0 Example Ref. No. R 1

-R

3 n p R 6 q R 17 B16 n-C 8

H

17 C0 0 0 CO 2 H 0 H 18 B17 n-C 7

H

15 C0 0 0 CO 2 H 0 H 19 B18 n-C 6

H

13 C0 0 0 CO 2 H 0 H B 19 n-C 13 H 27 C0 0 0 H 0 H 21 B20 n-C 9

H

19 C0 0 0 H 0 H 22 B21 n-C 1

-H

27 C0 1 0 H 0 H 23 B22 n-C 13

H

2 7 C0 2 0 H 0 H 24 B23 n-C 13

H

27 C0 4 0 H 0 H B24 n-C 1 3

H

27 C0 0 0 CONH, 0 H 26 B25 n-C 9 Hl 9 C0 0 0 CONH- 2 0 H 27 B26 n-C 1 3

H

27 C0 0 0 CO,Me 0 H 28 B27 n-C, 3

H

27 C0 0 0 H I COH 29 B28 n-C 9

H

19 C0 1 0 H 1 C0 2

H

For all Examples shown: X=Y0; R 4

=R

5 m=0; Rg=phosphono; R 9

=H.

*the stereochemnistry of the carbon atom to which R, is attached is S.

**the sereocemnsry oftecarbon atom to which R, is attached is R.

is C00 8 i H and Rg is phosphono. 200 WO 98/50399 PCT/US98/09385 14 EXAMPLE 1 Preparation of 3 -n-alkanoyloxytetradecanoic acids A solution of methyl 3-oxotetradecanoate (19 g, 0.074 mol) in MeOH (100 mL) was degassed by sparging with argon (15 min). [(R)-Ru(Binap)C1] 2 .NEt3 catalyst (0.187 g, 0.111 mmol) and 2 N aqueous HCI (0.5 mL) were added and the resulting mixture was hydrogenated at 60 psig and 40-50°C for 18 h. The reaction was diluted with hexanes (250 mL), filtered through a short column of silica gel, and concentrated. The crude product was dissolved in tetrahydrofuran (THF; 200 mL), treated 2.4 N aqueous LiOH (83 mL, 0.2 mol) and stirred vigorously at room temperature for 4 h. The resulting slurry was partitioned between ether (200 mL) and 1 N aqueous HCI (200 mL) and the layers separated. The aqueous layer was extracted with ether (100 mL) and the combined ethereal extracts were dried (Na 2

SO

4 and concentrated. The crude hydroxy acid was dissolved in hot acetonitrile (250 mL), treated with dicyclohexylamine (DCHA; 17 mL, 0.085 mol) and stirred at 60°C for 1 h. The product that crystallized upon cooling was collected and recrystallized from acetonitrile (650 mL) to yield 28.6 g ofdicyclohexylammonium (R)-3-hydroxytetradecanoate as a colorless solid: mp 94-95°C; 'H NMR (CDCI 3 6 0.88 3 H, J 6.5 Hz), 1.05-1.58 24 1.65 (m, 2 1.80 4 2.01 (br d, 4 H) 2.18 (dd, 1 H, J= 15.7, 9.4 Hz), 2.36 (dd, 1 H, J= 15.7, 2.6 Hz), 2.94 2 3.84 1 H) To a mixture of the compound prepared in above (50 g, 0.117 mol) and 2,4'-dibromoacetophenone (39 g, 0.14 mol) in EtOAc (2.3 L) was added triethylamine (19.6 mL, 0.14 mol) and the resulting solution was stirred for 18 h at room temperature.

The voluminous precipitate that formed was collected and triturated with warm EtOAc (3 x 400 mL). The combined triturates and filtrate were washed with 1 M aq. HC1, saturated aq. NaCl and dried (Na 2

SO

4 Volatiles were removed under reduced pressure and the crude product obtained was crystallized from EtOAc-hexanes to give 47.2 g of (R)-3-hydroxytetradecanoic acid p-bromophenacyl ester as a colorless solid: mp 109-109.5°C; 'HNMR(CDC 3 5 0.88 3 H,J- 6.5 Hz) 1.15-1.70 20 2.56 (dd, 1 H, J= 15.1, 9.1 Hz), 2.69 (dd, 1 H, J= 15.1, 2.9 Hz), 3.27 (br s, 1 4.12 1 5.31 1 H, J= 16.5 Hz), 5.42 1 H, J= 16.5 Hz), 7.65 2 H, J= 8.5 Hz), 7.78 2 H, J= 8.5 Hz).

WO 98/50399 PCT/US98/09385 A solution of the compound prepared in above (4.6 g, 10.4 mmol) in CH,C1 2 (50 mL) containing 4 -dimethylaminopyridine (0.12 g, 1.0 mmol) and pyridine mL, 62 mmol) was treated at room temperature with myristoyl chloride (3.1 mL, 11.4 mmol). After stirring for 5 h at room temperature MeOH (0.5 mL) was added, and the reaction mixture was concentrated. The residue was partitioned between Et20 (150 mL) and cold 10% aqueous HC1 (50 mL) and the layers separated. The ethereal layer was dried (Na 2

SO

4 and concentrated and the residue obtained was purified on a short pad of silica gel with 5% EtOAc-hexanes. The diester was dissolved in AcOH (42 mL) and treated with three equal portions of zinc dust g, 90 mmol) at 60 0 C over a 1 h period.

After an additional hour at 60 0 C, the cooled reaction mixture was sonicated (5 min), filtered through Celite® and concentrated. The residue was purified by flash chromatography on silica gel with 10% EtOAc-hexanes to give 4.17 g of tetradecanoyloxytetradecanoic acid as a colorless solid: mp 28-29°C; 'H NMR (CDCl 3 6 0.88 6 1.15-1.40 38 1.50-1.70 4 2.28 2 H, J= 7.4 Hz), 2.56 (dd, 1 H, J= 15.9, 5.8 Hz), 2.63 (dd, 1 H, J= 15.9, 7.1 Hz), 5.21 1 H).

In the same manner as described in Example the compound prepared in Example (2.5 g, 5.68 mmol) was acylated with lauroyl chloride (1.45 mL, 6.25 mmol) in the presence of pyridine (0.57 mL, 7.0 mmol) in CH 2 C2 (60 mL) and then deprotected with zinc (9.3 g, 142 mmol) in AcOH (40 mL) to afford dodecanoyloxytetradecanoic acid as a colorless oil: 'H NMR (CDC1 3 8 0.90 6 H, J 6.5 Hz), 1.0 1.75 46 2.30 2 2.62 2 5.22 1 H).

A solution of the compound prepared in Example (2.5 g, 5.68 mmol) was treated with undecanoic acid (1.16 g, 6.25 mmol) and EDCMel (2.08 g, 7.0 mmol) in CH 2 C1 2 (60 mL) and then deprotected as described in Example with zinc (9.3 g, 142 mmol) in AcOH (40 mL) to afford (R)-3-undecanoyloxytetradecanoic acid as a colorless oil: 'H NMR (CDC1 3 8 0.89 6 H, J= 6.7 Hz), 1.0 1.75 44 2.29 (m, 2 2.61 2 5.22 1 H).

In the same manner as described in Example the compound prepared in Example (4.4 g, 10 mmol) was acylated with decanoyl chloride (2.3 mL, 11 mmol) in the presence of pyridine (1.2 mL, 15.0 mmol) in CH 2

C

2 I (100 mL) and then deprotected with zinc (16.4 g, 250 mmol) in AcOH (60 mL) to afford WO 98/50399 WO 9850399PCT/US98/09385 16 decanoyloxytetradecanoic acid as a colorless oil: 1H NMR (CDCl 3 6 0.89 6 H, J 6.8.Hz), 1.0 1.75 (in, 34 2.29 2 H, J= 7.4 Hiz), 2.61 2 H, J= 4.2 Hz), 5.22 (in, I H).

In the same manner as described in Example the compound prepared in Example (2.5 g, 5.68 mmol) was acylated with nonanoyl chloride 13 mL, 6.25 mmol) in the presence of pyridine (0.57 mL, 7.0 inmol) in CH 2

CI

2 (60 mL) and then deprotected with zinc (9.3 g, 142 inmol) in AcOH (40 mL) to afford nonanoyloxytetradecanoic acid as a colorless oil: 'H NMR (CDCl 3 6 0.89 6 H, J 6.9 Hz), 1.0 1.75 (in, 32 2.29 2 H, J 7.5 Hz), 2.61 (mn, 2 5.22 (in, 1 H).

In the same manner as described in Example 1 the compound prepared in Example (2.5 g, 5.68 iniol) was acylated with octanoyl chloride (1.07 mL, 6.25 inmol) in the presence of pyridine (0.57 mL, 7.0 inmol) in CH 2 Cl 2 (60 inL) and then deprotected with zinc (9.3 g, 142 minol) in AcOH (40 mL) to afford octanoyloxytetradecanoic acid as a colorless oil: 'H NMR (CDC 3 )860.92 6 H, J= 6.9 Hz), 1.0 -1.75 (mn, 30 2.32 2 H, J= 7.4 Hz), 2.63 2 H, J= 4.4 Hz), 5.23 (mn, 1

H).

In the same manner as described in Example 1 the compound prepared in Example 1 (2.5 g, 5.68 inmol) was acylated with heptanoyl chloride (0.97 mL, 6.25 inmol) in the presence of pyridine (0.57 mL, 7.0 mmol) in CH 2 Cl 2 mL) and then deprotected with zinc (9.3 g, 142 minol) in AcOH (40 mL) to afford (R)-3-heptanoyloxytetradecanoic acid as a colorless oil: 'H NMR (CDCl 3 6 0.89 6 H, J= 6.8 Hz), 1.0 1.75 (in, 28 2.29 2 H, J 7.4 Hz), 2.61 2 H, J= 5.8 Hz), 5.22 (in, 1 H).

EXAMPLE 2 (B I) Preparation of 3 -Hydroxy-(S)-2-[(R)-3 -tetradecanoyloxytetradecanoylamino]propyl 2- Deoxy-4- O-phosphono-2 3 -tetradecanoyloxytetradecanoylIamino] -3 0- 3 tetradecanoyloxytetradecanoyl] -1-D-gl ucopyranoside TriethylIamnmoniumn Salt (Compound R,=R 2

=R

3 3

H

2 7 C0, X=Y=O, n=in=q=0, R 4

=R

5

=R

7 =Rq=H, R6=OH, p=1, R 8 =P0 3

H

2 To a solution of 2-(triinethylsilyl)ethyl 2-amnino-2-deoxy-4,6-0isopropylidene-p3-D-glucopyranoside (6.46 g, 20.2 nimol) in CHC1 3 (300 mL) was added WO 98/50399 PCT/US98/09385 17 1 N aqueous NaHCO 3 (300 mL) and 2 ,2,2-trichloroethyl chloroformate (8.5 g, 40 mmol).

The resulting mixture was stirred vigorously for 3 h at room temperature. The organic layer was separated, dried (Na2SO 4 and concentrated to give a colorless syrup. Flash chromatography on silica gel (gradient elution, 30-40% EtOAc-hexanes) afforded 9.6 g of 2-(trimethylsilyl)ethyl 2-deoxy-4,6-O-isopropylidine-2-(2,2,2trichloroethoxycarbonylamino)-p-D-glucopyranoside as a colorless solid: mp 69-70°C; 'H NMR (CDCl 3 8 0.0 9 0.94 2 1.44 and 1.52 (2s, 6 2.94 (br s, 1 H), 3.23-3.37 2 3.48-3.62 2 3.79 1 H, J= -10.5 Hz), 3.88-4.08 3 H), 4.65 1 H, J=8.3 Hz), 4.74 2 5.39 1 H, J=7.4 Hz).

A solution of the compound prepared in above (7.5 g, 15.2 mmol), 3 -tetradecanoyloxytetradecanoic acid (7.58 g, 16.7 mmol) and 4 -pyrrolidinopyridine (0.25 g, 1.7 mmol) in CH 2 C1 2 (95 mL) was treated with 1-(3-dimethylaminopropyl)-3ethylcarbodiimide methiodide (EDC-Mel; 4.94 g, 16.7 mmol) and stirred for 16 h at room temperature. The reaction mixture was filtered through a short pad of Celite®, concentrated, and the resulting residue was heated at 60°C in 90% aqueous AcOH (100 mL) for 1 h. The mixture was concentrated and residual AcOH and water were removed by azeotroping with toluene (2 x 150 mL). The crude diol was purified by flash chromatography on silica gel (gradient elution, 30-40% EtOAc-hexanes) to give 11.8 g of2-(trimethylsilyl)ethyl 2 -deoxy- 3 -O-[(R)-3-tetradecanoyloxytetradecanoyl]-2- 2 2 2 -trichloroethoxycarbonylamino)-P-D-glucopyranoside as an amorphous solid: 'H NMR (CDCl 3 80.0 9 0.9 8 1.1-1.7 42 2.30 2 H, J=7.4 Hz), 2.52 2 3.36-3.72 4 3.78-4.03 3 4.57 1 H, .J=8.3 Hz), 4.65 (d, 1 H, J=11 Hz), 4.77 1 H, J=11 Hz), 5.0-5.15 2 5.20 1 H, J=7.4 Hz).

A solution of the compound prepared in above (10.9 g, 12 mmol) and pyridine (2 mL, 25 mmol) in CH2C1 2 (125 mL) at 0 C was treated dropwise over 15 min with a solution of 2,2,2-trichloro-1,1-dimethylethyl chloroformate (3.17 g, 13.2 mmol) in CH 2 C1 2 (25 mL). The reaction mixture was allowed to warm slowly to ambient temperature over 3.5 h. 4-Pyrrolidinopyridine (0.89 g, 6.0 mmol), N,Ndiisopropylethylamine (10.5 mL, 60 mmol) and diphenyl chlorophosphate (3.7 mL, 18 mmol) were added sequentially and the resulting mixture was stirred for 5 h at room temperature. The reaction mixture was diluted with CHC1 2 (500 mL), washed with cold WO 98/50399 WO 9850399PCTIUS98/09385 18 aqueous HC1 (2 x 250 mL), water (250 mL), saturated aqueous NaHCO 3 (250 mL4, dried (Na 2

SO

4 and then concentrated. The residue obtained was purified by flash chromatography on silica gel eluting with 12.5% EtOAc-hexanes to give 15.1 g of 2-(trimethylsilyl)ethyl 2-deoxy-4-O-diphenylphosphono3 tetradecanoyloxytetradecanoyl] ,2-trichloro- 1,1-dimethylethoxycarbonyl)-2 2 2 2 -trichlorethoxycarbonylamino)- 3-D-glucopyrano side as a viscous oil: 'H NMR (CDC1 3 8 0.0 9 0.8-1.0 (mn, 8 1.1-1.65 (in, 42 1.83 and 1.90 (2s, 6 H), 2.15-2.45 4H), 3.34 I H,J= -8 Hz), 3.37 1 3.81 (in, 1 3.95 1 H), 4.27 (dd, I H, J= 12, 5Hz), 4.34 1 H, J=12 Hz), 4.5 8(d, I H, J=12 Hz), 4.66 1 H, J= 9Hz), 4.86 I H, J= 12 Hz), 5.03 1 H, J=7.9 Hz), 5.21 (in, 1 5.54-5.70 (in, 2 7.2-7.8 (mn, 10 H).

A solution of the compound prepared in above (1.87 g, 1.41 mmol) in

CH

2 Cl 2 (3 mL) at 0 0 C was treated dropwise over 10 min with trifluoroacetic acid (TFA; 6 mL) and then stirred for 4 h at 0 0 C. The reaction mixture was concentrated and residual TFA was removed by azeotroping with toluene (2 x 5 mL). A solution of the lactol and dimethylformamide (2.2 mL, 28.2 inmol) in CH 2 Cl 2 (14 mL) at 0 0 C was treated with oxalyl bromide (2.0 M in CH 2 C1 2 2.1 inL, 4.2 inmol) dropwise over 15 min and the resulting suspension was stirred at 0 0 C for 24 h. The reaction mixture was partitioned between cold saturated aqueous NaHCO 3 (25 mL) and ether (50 mL) and the layers were separated. The ethereal layer was washed with saturated aqueous NaCl, dried (Na-S0 4 and concentrated to give 1.85 g 100%) of 2-deoxy-4-O-,diphenylphosphono-3-o-[(R)- 3 -tetradecanoyloxytetradecanoyl]-6-o-(2,2,2-trichloro-1,1I -dimethylethoxycarbonyl-2- 2 2 2 -trichloroethoxycarbonylamino)-a-D-glucopyranosyI bromide as a colorless glass.

A solution of (R)-2-ainino-3-benzyloxy- I-propanol (0.46 g, 2.33 nimol) and (R)-3-tetradecanoyloxytetradecanoic acid (1.29 g, 2.83 inmol) in CH 2

CI

2 (20 mL) was treated with EDC&MeI (0.78 g, 2.79 n'mol) and stirred for 16 h at room temperature.

The reaction mixture was filtered through a short pad of Celites and concentrated. Flash chromatography on silica gel with 45% EtOAc-hexanes afforded 1.1 g of 3benzyloxy-(R)-2-[(R)-3 -tetradecanoyloxytetradecanoylamino]propanoI as a colorless solid: mp, 42-44.5 0 C; 'H NMR 8 0.88 6 H, J Hz), 1.0-1.7 (in, 42 2.50 2 WO 98/50399 PCT/US98/09385 19 H, J=7.5 Hz), 2.46 2 3.56 (br s, 1 3.5-3.75 3 3.78 (dd, 1 H, J=l 1, 4 Hz) 4.08 1 4.51 2 5.17 1 6.36 1 H, J=7.8 Hz), 7.2-7.4 5 H).

To a solution of the compound prepared in above (1.00 g, 0.776 mmol) and the compound prepared in above (0.35 g, 0.57 mmol) in dichloroethane (4.5 mL) was added powdered 4 A molecular sieves (1.25 g) and calcium sulfate (2.7 g, 20 mmol).

After stirring for 10 min at room temperature, the mixture was treated with mercury cyanide (1.0 g, 4.0 mmol) and then heated to reflux for 12 h shielded from light. The reaction mixture was diluted with CH 2

CL

2 (25 mL) and filtered through a pad of Celite®.

The filtrate was washed with 1 N aqueous KI (25 mL), dried (Na 2

SO

4 and concentrated.

The residue was chromatographed on silica gel with EtOAc-hexanes-MeOH (80:20:0-70:30:1, gradient elution) to give 0.66 g of 3-benzyloxy-(S)-2-[(R)-3tetradecanoyloxytetradecanoylamino]propyl 2-deoxy-4-O-phosphono-3-O-[(R)tetradecanoyloxytetradecanoyl]-6-0-(2,2,2-trichloro- 1,1-dimethylethoxycarbonyl)-2- 2 2 2 -trichloroethoxycarbonylamino)-p-D-glucopyranoside as an amorphous solid: 'H NMR 8 0.88 12 H, J= -6.5 Hz), 1.0-1.65 84 1.79 and 1.86 (2s, 6 2.1-2.5 8 3.35-3.55 3 3.65-3.8 3 4.1-4.75 9 5.05-5.3 2 5.3- 2 6.04 1 H, J=8.4 Hz), 7.05-7.45 15 H).

A stirred solution of the compound prepared in above (0.60 g, 0.328 mmol) in AcOH (9 mL) at 55 C was treated with zinc dust (1.1 g, 16 mmol) in three equal portions over 1 h. The cooled reaction mixture was sonicated, filtered through a bed of Celite® and concentrated. The resulting residue was partitioned between CH 2 Cl 2 mL) and cold 1 N aqueous HC1 (35 mL) and the layers separated. The organic layer was washed with 5% aqueous NaHCO 3 dried (Na 2

SO

4 and concentrated. A mixture of the residue obtained and (R)-3-tetradecanoyloxytetradecanoic acid (0.18 g, 0.39 mmol) in CH 2 Cl 2 (3.5 mL) was stirred with powdered 4A molecular sieves (0.1 g) for 30 min at room temperature and then treated with 2-ethoxy- 1 -ethoxycarbonyl- 1,2-dihydroquinoline (EEDQ; 0.12 g, 0.49 mmol). The resulting mixture was stirred for 6 h at room temperature, filtered through Celite® and then concentrated. Chromatography on silica gel (gradient elution, 0.5 1% MeOH-CHCl 3 afforded 0.31 g of 3-benzyloxy-(S)- 2 3 -tetradecanoyloxytetradecanoylamino]propyl 2-deoxy-4-O-diphenylphosphono- 2-[(R)-3-tetradecanoyloxytetradecanoylamino]-3-0-[(R)-3- WO 98/50399 PCT/US98/09385 tetradecanoyloxytetradecanoyl]--D-glucopyranoside as an amorphous solid: 'H NMR

(CDC

3 8 0.88 18 H,J= -6.5 Hz), 1.0-1.8 126 2.1-2.5 12 3.35-3.75 6 3.80 2 4.23 1 4.46 1 H, J=12 Hz), 4.51 1 H, J=12 Hz), 4.65 1 H, J= -9.5 Hz), 4.82 1 H, J=8.1 Hz), 5.05-5.25 3 5.47 1 H, J= -9.5 Hz), 6.16 1 H, J=8.1 Hz), 6.31 1 H, J=8.4 Hz), 7.1-7.4 15 H).

A solution of the compound prepared in above (0.26 g, 0.138 mmol) in THF (25 mL) was hydrogenated in the presence of 5% palladium on carbon (50 mg) at room temperature and atmospheric pressure for 16 h. After removal of the catalyst by filtration, AcOH (3 mL) and platinum oxide (0.14 g) were added and the hydrogenation was continued at room temperature and 75 psig for 24 h. The resulting opalescent reaction mixture was diluted with 2:1 CHCl 3 -MeOH (20 mL) and sonicated briefly to give a clear solution. The catalyst was collected, washed with 2:1 CHC1 3 -MeOH (2 x mL) and the combined filtrate and washings were concentrated. The residue was dissolved in 1% aqueous triethylamine (10 mL) by sonicating for 5 min at 35°C and the resulting solution was lyophilized. Flash chromatography on silica gel with chloroformmethanol-water-triethylamine (94:6:0.5:0.5-88:12:1.0:1.0, gradient elution) afforded 0.20 g of product as a colorless powder. A portion of the chromatography product (0.166 g) was dissolved in cold 2:1 CHC13-MeOH (33 mL) and washed with cold 0.1 N aqueous HCI (14 mL). The lower organic layer was filtered and concentrated and the free acid obtained was lyophilized from 1% aqueous triethylamine (pyrogen free, 15 mL) to give 0.160 g of 3 -hydroxy-(S)-2-[(R)-tetradecanoyloxytetradecanoylamino]propyl 2deoxy-4-O-phosphono-2-[(R)-3-tetradecanoyloxytetradecanoylamino]-30- tetradecanoyloxytetradecanoyl]--D-glucopyranoside triethylammonium salt as a colorless solid: mp 178-180 0 C (dec); IR (film) 3293, 3103, 2959, 2924, 2855, 1732, 1654, 1640, 1553, 1467, 1377, 1259, 1175, 1106, 1086, 1050, 803, 720 HMR (CDCl 3 -CD30D) 8 0.88 18 H,J= -7 Hz), 1.0-1.7 135 2.15-2.75 12 H), 3.02 6 H, J=7 Hz), 3.35-4.1 7 4.22 1 H, J 9.5 Hz), 4.77 1 H, J=8 Hz), 5.05-5.35 4 6.58 1 H, J= 6 Hz), 6.73 1 H, J= 7.5 Hz, NH); 3 C NMR (CDCl1) 8 173.5, 173.2, 170.7, 170.5, 170.0, 100.7, 75.9, 72.7, 71.2, 71.0, 70.8, 70.6, 67.9, 61.7, 60.5, 55.0, 50.4, 45.6,41.4, 39.5, 34.5,34.4,32.0,31.8,30.3,29.8,29.4,29.3, 25.3, 25.1, 22.7, 14.2, 8.6.

WO 98/50399 WO 9850399PCTIUS98/09385 21 Anal. Calcd for C 19

H,

92

N

3 0 18 P -5 H 2 0 C, 64.84; H, 11. 10; N, 2.29; P, 1. 69.

Found: C, 64.69; H, 11.24; N, 1.93; P, 1.44.

EXAMPLE 3 (B32) Preparation of 3 -Hydroxy-(R)-2-[(R)-3 -tetradecanoyloxytetradecanoylamino]propyl 2- Deoxy-4-O-phosphono-2- -tetradecanoyloxytetradecanoylamino]-3 tetradecanoyloxytetradecanoyl] 3-D-gl ucopyrano side Triethylammonium Salt (Compound R,=R,=R 3 n-C 3

H

27 C0, X=Y=O, n=m=q=0, R,=R 5

=R

7

=R

9 R6=01H, p=l, R 8 =P0 3

H

2 A solution of the compound prepared in Example (0.63 g, 1.02 mmol) in CH 2 C1 2 (7 mL) was treated sequentially with pyridine (0.4 mL, 5 mmol), 4dimethylaminopyridine (cat.) and 2,2,2-trichioro- 1,1 -dimethylethyl chloroformate (0.307 g, 1.23 mmol) and stirred for 16 h at room temperature. The reaction mixture was diluted with CH,Cl, (25 mL), washed with saturated aqueous NaHCO 3 (25 mL) and dried (Na 2

SO

4 Removal of volatiles in vacuo gave a residue which was dissolved in THF- AcOH (10 mL, 9: 1) and hydrogenated in the presence of 5% palladium on carbon (150 mg) at room temperature and atmospheric pressure for 24 h. After removal of the catalyst by filtration and concentration of the filtrate, the residue was purified by flash chromatography on silica gel with 35% EtOAc-hexanes to give 0.536 g of 3- 2 2 2 -t r ich Io ro -I,I -d im eth y Ieth ox y carb on yIo xy) 2 -3 tetradecanoyloxytetradecanoylamino]propanol as an amorphous solid: 'H NMR (CDCl 3 8 0.88 6 H, J= -6.5 Hz), 1. 1-1.7 (in, 42 1.94 6 2.30 2 H, J=7.5 Hz), 2.47 2 H, J=6 Hz), 3.50 (br s, I 3.72 (in, 2 4.15-4.35 (in, 3 5.15 (in, 1 6.18 1 H, J=7.2 Hz).

In the same manner as described in Example the compound prepared in above (0.3 10 g, 0.426 mmol) and the compound prepared in Example 2- (0.961 g, 0.745 inmol) were coupled in the presence of mercury cyanide (0.43 g, 1.7 inmol) to give 0.644 g of 3-(2,2,2-trichloro- 1,1 -dimethylethyloxycarbonyloxy)- 2 3 -tetradecanoyloxytetradecanoylamino]propyl 2 -deoxy-4-0-phosphono-3 -0- [(R)-tetradecanoyloxytetadecanoyl] -6.0..(2,22tichloro- 1, 1 -dimethylethoxycarbonyl)- 2 2 2 2 -trichloroethoxycarbonyamino)-p3-D-glucopyranoside as an amorphous solid: 1H NMR (CDCl 3 )8 0.88 12 H, J= -6.5 Hz), 1.0-1 .7 (mn, 84 1.81 and 1.89 (2s, 6 H), WO 98/50399 WO 9850399PCTIUS98/09385 22 1.93 6 2.15-2.55 (in, 8 3.45-3.7 (in, 2 3.80 (br d, I H, J=9 Hz), 3.9-4.45 6 4.6-4.8 (in, 3 4.87 1 H, J=8.1 Hz), 5.0-5.25 (in, 2 5.48 1 H, J Hz), 6.1-6.3 (in, 2 H).

In the same manner as described in Example the compound prepared in above (0.602 g, 0.3 10 inmol) was deprotected with zinc (1.5 g, 23 iniol) and acylated with (R)-3-tetradecanoyloxytetradecanoic acid, 17 g, 0.3 7 inmol) in the presence of EEDQ 115 g, 0.467 mmol) to give 0.3 65 g of 3-hydroxy-(R)-2-[(R)- 3 -tetradecanoyloxytetradecanoylamino]propyI 2-deoxy-4-O-diphenylphosphono-24(R).

3 -tetradecanoyloxytetradecanoyamino-3..o4(R).3-tetradecanoyloxytetradecanoyl]-3Dglucopyranoside as an amorphous solid: 'H NMR (CDCI 3 8 0.88 18 H, J= 6.5 Hz), 1.0-1.7 (in, 126 2.15-2.55 (mn, 12 3.18 (br s, 1 3.45-3.8 (in, 8 3.85-4.05 (in, 2 4.69 I H, J= -9.5 Hz), 5.05-5.25 (in, 3 5.42 1 H, J=-9.5 Hz), 6.42 1 H, J=7.8 Hz), 6.5 9 I H, J=7.2 Hz), 7.1-7.4 (in, 10 H).

In the same manner as described in Example the compound prepared in above (0.355 g, 0.196 inmol) was hydrogenated in the presence of platinum oxide (175 ing) to give 0.265 g of 3-hydroxy-(R)-2-[(R)-3tetradecanoyloxytetradecanoylamino]propyl 2 -deoxy-4-O-phosphono-2- tetradecanoyloxytetradecanoylamino-3O[(R)3tetradecanoyloxytetradecanoyl]-oDglucopyranoside triethylanimoniuin salt as a colorless solid: mp 159-1 60'C; JR (film) 3291, 2956, 2922, 2853, 1738, 1732, 1716, 1650, 1643, 1556, 1468, 1171, 1109, 1083, 1051 11H1NMR (CDC 3

-CD

3 OD) 850.88 18 H,J= -6.5 Hz), 1.0-1.7 (in, 135 H), 2.15-2.75 (in, 12 3.06 6 H, J=7 Hz), 3.25-3.45 (in, 2 3.5-4.05 (in, 12 4.19 1 H, J -9.5 Hz), 4.48 I H, J=8.4 Hz), 5.04-5.26 (in, 4 7.18 1 H, J=7.8 Hz), 7.27 1 H, J=8.7 Hz); 1 3 C NMR (CDC 3 6 173.5, 173.4, 170.7, 170.6, 170. 1, 101.0, 76.0, 72.6, 71.4, 71.0, 70.8, 70.6, 68.7, 61.8, 60.5, 55.3, 50.5, 45.6, 41.5, 41.4, 39.5, 34.6, 34.4, 34.3, 32.0, 29.8, 29.4, 25.4, 25.1, 22.7, 14.1, 8.6.

Anal. Calcd for C 99

H,

92

N

3 0, 8 P H,0: C, 67.50; H, 11.10; N, 2.39; P, 1.76.

Found: C, 67.40; H, 11.22; N, 2.34; P, 2.11.

WO 98/50399 PCT/US98/09385 23 EXAMPLE 4 (B3) Preparation of 3-Hydroxy-(S)-2-[(R)- 3 -dodecanoyloxytetradecanoylamino]propyl 2- Deoxy-4-O-phosphono-2-[(R)-3-dodecanoyloxytetradecanoylamino]-3-0-[(R)-3dodecanoyloxytetradecanoyl]--D-glucopyranoside Triethylammonium Salt (Compound

R=R

2

=R

3 n-C,,H 2 3 CO, X=Y=O, n=m=q=0, R 4 R=R=R=R=H, R6=OH, p=l, Rs=PO 3

H

2 A solution of D-glucosamine hydrochloride (20 g, 92.8 mmol) in H,O (250 mL) was treated with a saturated aqueous NaHCO 3 (250 mL) and 2,2,2trichloroethyl chloroformate (14.05 mL, 102 mmol) and stirred vigorously for 18 h. The white solid that formed was collected on a fritted funnel and dried under vacuum for 24 h. A solution of the solid in pyridine (100 mL) was cooled to 0 oC and treated with acetic anhydride (100 mL) via addition funnel. The solution was stirred for 18 h at room temperature, poured into 1 L of H 2 0 and extracted with CHC1 3 (3 x 500 mL). The solvent was removed in vacuo to afford 45 g (quant.) of N-(2,2,2trichloroethoxycarbonylamino)-1,3,4,6-tetra-O-acetyl-2-deoxy-a-D-glucopyranoside which was used without furthur purification: 'H NMR (CDCl 3 5 2.06 6 2.12 (s, 3 2.22 3 4.03 1 4.07 1 H, J= 12.4 Hz), 4.22 (dt, 1 H, J= 9.9, 3.6 Hz), 4.30 (dd, 1 H, J= 12.4, 4.0 Hz), 4.64 1 H, J= 9.6 Hz), 5.28 (dt, 1 H, J= 10.2, 9.9 Hz), 6.25 1 H, J= 3.6 Hz).

A solution of (R)-2-amino-3-benzyloxy- -propanol (5 g, 27.6 mmol) in

CH

2

CI

2 (250 mL) was treated with allyl chloroformate (3.2 mL, 30 mmol) and saturated aqueous NaHCO 3 (250 mL) for 18 h. The organic layer was separated and concentrated in vacuo. Purification by chromatography eluting with 30 EtOAc hexanes afforded 6.9 g (94 of (R)-2-(allyloxycarbonylamino)-3-benzyloxy-1 -propanol as an amorphous solid: 'H NMR (CDC1 3 6 2.56 (br s, 1 3.69 3 3 88 2 4.54 2 H), 4.58 2 H, J= 5.6 Hz), 5.23 (dd, 1 H, J= 10.4, 1.1 Hz), 5.33 (dd, 1 H,J= 17.1, 1.1 Hz), 5.42 1 5.93 1 7.35 5 H).

A solution of the compounds prepared in and above (8.9 g, 17 mmol and 3.6 g, 10 mmol, respectively) in CH2C1 2 was treated with boron trifluoride etherate (4.3 mL, 34 mmol) at room temperature for 16 h. The reaction mixture was quenched with saturated aq. NaHCO 3 (100 mL) and extracted with EtOAc (3 x 100 mL).

The combined EtOAc extracts were dreid (Na 2

SO

4 and concentrated. The residue WO 98/50399 WO 9850399PCTIUS98/09385 24 obtained was chromatographed with 20 EtOAc hexanes to afford 6.03 g (83 of 3 -benzyloxy-(S)-2-(allyloxycarbonylamino)propyl 2-deoxy-3,4,6-tri-O-acetyl-2-(2,2,2.

trichloroethoxycarbonylamino)-p3-D-glucopyranoside as an amorphous solid: 'H NMR (CDCl 3 8 2.02 3 2.03 3 2.08 3 3.45 (in, 1 3.54 (in, I 3.64 (in, 1 3.76 1 H,J =7.2 Hz), 3.91 (mn,2 4.12 1 H, J= 12.2 Hz), 4.26 (dd, 1 H, J 12.4, 4.7 Hz), 4 .3 7 I H, J =8.2 Hz), 4.43 1 H, J 12.1 Hz), 4.5 5 (in, 2 4.68 (in, 2 4.87 I H, J= 8.0 Hz), 5.07 (in, 2 5.21 1 H, J= 9.7 Hz), 5.29 1 H,J 17.3 Hz), 5.91 (in, 1 7.3 6 (in, 5 A solution of the compound prepared in above (6.0 g, 8.3 mmol) in methanol (83 mL) was treated with ammonium hydroxide (8.3 mL) at room temperature for 2 h. The solvent was removed in vacuo and replaced with 2,2-dimethoxypropane mL) and cainphorsulfonic acid (100 mg) was added. The reaction was stirred for 18 h, neutralized with solid NaHCO 3 (1 filtered and concentrated in vacuo. Purification by chromatography with 50 EtOAc hexanes afforded 4.58 g (86 of 3-benzyloxy-(S)- 2 -(allylIoxycarbony lamino)propylI 2-deoxy-4,6-O-isopropylidine-2-(2,2,2trichloroethoxycarbonylanino)-p3-D-glucopyranoside: 'H NMR (CDCl 3 6 1.46 3 H), 1.53 3 2.94 (in, 1 3.25 (in, 1 3.55 (in, 4 3.83 (in, 3 3.93 (in, 3 H), 4.52 (mn, 5 4.68 I H, J 12.1 Hz), 4.77 1 H, J 12.1 Hz), 5.07 (in, 1 5.26 (in, 2 5.92 (in, 1 7.37 (in, 5 H).

A solution of the compound prepared in above (1.0 g, 1.56 iniol) in

CH

2 C1 2 (20 inL) was treated with (R)-3-dodecanoyloxytetradecanoic acid (730 mg, 1.71 inmol) in the presence of EDC-MeI (560 mg, 1.87 mmol) and 4-pyrrolidinopyridine ing). The reaction was stirred at room temperature for 18 h and filtered through a 6 x 8 cm plug of silica gel using 20 EtOAc hexanes as eluent to afford 1.33 g (82 of 3benzyloxy-(S)-2-(allyloxycarbonylamnino)propyl 2-deoxy-4,6-O-isopropylidene-3-O-[(R)- 3 -do decanoyl oxytetradecanoy] -2 tri chi oroethoxycarbony amino)- 0 D glucopyranoside as an amorphous solid: 'H NMR (CDCl 3 50.88 6H, J=6.8 Hz), 1.1I 1.6 (in, 38 1.37 3 1.46 3 2.28 2 H, J= 7.4 Hz), 2.49 (dd, 1 H, J= 15.1, 6.0 Hz), 2.61 (dd, 1H,J 15.1, 6.6 Hz), 3.25 -4.0(in, 9H), 4.38(in, 2H), 4.54 (in, 2 4.65 (in, 2 4.97 (in, 2 5.25 (in, 5 5.88 (in, 1 7.34 (in, 5 H).

WO 98/50399 PCT/US98/09385 To a solution of the compound prepared in above (1.31 g, 1.25 mmol) in THF (20 mL) was added dimethyl malonate (1.0 mL, 0.88 mmol) and the solution was degassed in a stream of argon for 30 min. Tetrakis(triphenylphosphine)palladium(0) (200 mg) was added and the reaction was stirred at room temperature for 2 h, and then concentrated in vacuo. The residue obtained was chromatographed on silica gel eluting with 5 -10% EtOAc CHC1 3 The free amine obtained was acylated with dodecanoyloxytetradecanoic acid (560 mg, 1.38 mmol) in the presence of EEDQ (370 mg, 1.5 mmol) in CH 2 C1 2 (15 mL). After stirring at room temperature for 18 h, the solvent was removed in vacuo and the resultant oil was chromatographed on silica gel eluting with 20 %EtOAc hexanes to afford 1.02 g (63 of 3-benzyloxy-(S)-2-[(R)-3dodecanoyloxytetradecanoylamino]propyl 2 -deoxy-4,6-O-isopropylidene-3-O-[(R)-3dodecanoyloxytetradecanoyl]-2-(2,2,2-trichloroethoxycarbonylamino)-p-Dglucopyranoside as a colorless amorphous solid: 'H NMR (CDCl 3 6 0.88 12 H, J= 6.9 Hz), 1.1 1.7 78 1.38 3 1.46 3 2.26 4 2.49 (dd, 1 H, J 15.1, 6.0 Hz), 2.61 (dd, 1 H,J= 15.1, 6.6 Hz), 3.25 4.0 9 5.01 2 6.02 1 H, J= 8.4 Hz), 7.34 5 H).

The compound prepared in above (1.0 g, 0.78 mmol) was treated with aqueous AcOH (20 mL) for 1 h at 60 The solution was concentrated in vacuo and residual AcOH and H 2 0 were removed by azeotroping with toluene (10 mL). The residue was dissolved in CHC1 2 cooled to 0 and treated with pyridine (0.076 mL, 0.94 mmol) and a solution of 2,2,2-trichloro-1,1-dimethylethyl chloroformate (205 mg, 0.86 mmol) in CH 2 C1 2 (5 mL). The reaction mixture was then allowed to warm and stir at room temperature for 18 h. The resulting light yellow solution was treated with diphenyl chlorophosphate (0.24 mL, 1.17 mmol), triethylamine (0.22 mL, 1.56 mmol) and catalytic 4-pyrrolidinopyridine (50 mg), and then stirred an additional 24 h at room temperature. The reaction mixture was diluted with Et20 (100 mL) and washed with aq. HC1 (50 mL). The organic phase was separated, dried over Na 2

SO

4 and concentrated in vacuo. Chromatography over silica gel using 10 EtOAc hexanes afforded 1.13 g (85 of 3-benzyloxy-(S)-2-[(R)-3dodecanoyloxytetradecanoylamino]propyl 2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3dodecanoyloxytetradecanoyl]-6-0-(2,2,2-trichloro-1,1-dimethylethoxycarbonyl)-2-(2,2,2- WO 98/50399 WO 9850399PCTIUS98/09385 26 trichloroethoxycarbonylamino)-I3-D-glucopyranoside as a colorless amorphous solid: 11H1NMR (CDCl 3 860.87 12 H, J= 6.9 Hz), 1. 1 1.6 (in, 78 1.78 3 1.86 (s, 3 2.01 (in, I 2.18 (in, 3 2.40 (in, 2 2.67 (in, I 2.88 1 H, J1 6.6 Hz), 2.97 (d,1 H, J =6.9 Hz), 3.41 (in, 2 3.72 (in, 1 3.82 (in, 1 4.24 (in, I H), 4.42 I H, J 11. 8Hz), 4.64 (in, 3 5.16 (in, 1 5.3 9 (in, 2 5.75 1 H, J= 4.3 Hz), 6.05 1 H, J= 8.4 Hz), 7.23 (in, 15 H).

In the same manner as described in Example the compound prepared in above (1.1 g, 0.65 inmol) was deprotected with zinc (2.1 g, 32 mmol) and acylated with (R)-3-dodecanoyloxytetradecanoic acid (330 mng, 0.78 inmol) in the presence of EEDQ (230 mg, 0.94 minol) to afford 399 mg (37 of 3-benzyloxy-(S)-2- 3 -dodecanoyloxytetradecanoylamino]propyI 2 -deoxy-4-O-diphenylphosphono-2- [(R)-3-dodecanoyloxytetradecanoylainino]-3 -dodecanoyltetradecanoyl]-3-Dglucopyranoside as a colorless amorphous solid.

In the same manner as described in Example the compound prepared in above (399 mg, 0.24 inmol) was hydrogenated in the presence of palladium hydroxide (15 0mg) on carbon in EtOH (10 mL) and platinum oxide (3 00 ing) in EtOH AcOH (10:1) to afford 65 mng (16 of 3-hydroxy-(S)-2-[(R)-3dodecanoyloxytetradecanoylainino]propyl 2 -deoxy-4-O-phosphono-2- dodecanoyloxytetradecanoylamino)-3 -dodecanoyloxytetradecanoy]-p-.D.

glucopyranoside triethylammoniuin salt as a white powder: mp 181 1840 C (dec): JR (filin)3306,2956,2922,2852, 1732,1644,1549,1467,1377,1164,1106,1051,721 cm- HNMR (CDC1 3

CD

3 OD) 850.88 18 H, J= 6.7 Hz), 1. 1 1.7 (in, 123 2.2 -2.7 (in, 12 3.06 6 H, J= 7.1 Hz), 3.3 4.0 (in, 13 4.23 (in, 1 4.44 1 H,J 7.7 Hz), 5.0 5.3 (in, 4 1 3 C NMR (CDCl 3 8 173.9, 173.5, 173.3, 170.8, 170.5, 170. 1, 101.0, 75.5, 73.0, 71.1, 70.9, 70.6, 67.9, 61.6, 60.7, 54.4, 50.4, 45.8, 41.6, 41.4, 39.6, 34.6, 31.9, 29.7, 29.4, 29.3, 25.4, 25.1, 22.7, 14.2,8.6.

Anal. Calcd. for C 93

H-

180

N

3 0 18 P C, 66.59; H, 10.94; N, 2.50; P, 1.85.

Found: C, 66.79; H, 10.65; N, 2.36; P, 1.70.

WO 98/50399 PCT[US98/09385 27 EXAMPLE 5 (B34) Preparation of 3 -Hydroxy-(S)-2-[(R)-3-undecanoyloxytetradecanoylamino]propyl 2- Deoxy-4-0-phosphono-2-[(R)-3 -undecanovloxytetradecanoylamino] undecanoyloxytetradecanoyl]-3-D-glucopyranoside Triethylammonium Salt (Compound

R

1

=R

2

=R

3 n-C 10

H

21 C0, X=Y=O, n=m=q=0, R 4 =Rs=R 7

=R

9 R6=01H, p=l,

R

8 =P0 3

H

2 In the same manner as described in Example the compound prepared in Example (1.0 g, 1.56 mmol) was acylated with undecanoyloxytetradecanoic acid (705 mg, 1.71 mmol) in the presence of EDC-MeI (560 mg, 1.87 mmol) and 4 -pyrrolidinopyridine (50 mg) in CH 2 Cl, (20 mL) to afford 1.23 g (77 of 3 -benzyloxy-(S)-2-(allyloxycarbonylamino)propyI 2-deoxy-4 sopropylIidene- 3 0- 3 -undecano yloxytetradecanoyI 2 ,2,2 trichloroethoxycarbonylamino)-3D-glucopyranoside as an amorphous solid: 1 1- NMR (CDCl 3 860.88 6 H, 6.9 Hz), 1. 1 1.6 (in, 36 1.3 7 3 1.46 3 2.28 2H), 2.52 (dd, 1 H, J= 15.1, 6.0OHz), 2.61 (dd,1I H, 15.5, 6.8 Hz), 3.25 1 H), 3.35 4.0 (in, 9 4.31 (mn, 2 4.54 (mn, 2 4.64 (mn, 2 5.02 (mn, 2 5.18 (mn, 2 5.25 (in, 1 5.86 (mn, I 7.34 (in, 5 H).

In the same manner as described in Example the compound prepared in above (1.21 g, 1. 17 inmol) was deprotected in THF (20 inL) in the presence of diinethyl inalonate (1.0 mL, 0.88 mmol) and tetrakis(triphenylphosphine)palladium(O) (200 mng) and then acylated with (R)-3-undecanoyloxytetradecanoic acid (540 mng, 1.30 inmol) in the presence of EEDQ (370 mg, 1.5 rmol) to afford 921 mng (61 of 3benzyloxy-(S)-2- -undecanoyloxytetradecanoylainino]propyI 2-deoxy-4,6-0isopropylidene-3 -undecanoyloxytetradecanoyl]2(2,22.

trichloroethoxycarbonylamino)-p-Dglucopyranoside as a colorless amorphous solid: 1H NMR (CDCl 3 80.88 12 H, J=6.6 Hz), 1.1 1.7 72 1.38 3 1.46 (s, 3 2.26 (in, 3 2.3 8 (mn, 5 2.49 (dd, 1 H, .1=15.2, 6. 0 Hz), 2.61 (dd, 1 H,J 15.0, 6.5 Hz), 3.25 4.0 (in, 9 4.30 (in, 2 4.59 (in, 3 6.03 I H,J= 8.2 Hz), 7.34 (in, 5 H).

In the same manner as described in Example the compound prepared in above (910 g, 0.71 inmol) was deprotected in 90 aqueous AcOH (20 mL), and then treated with pyridine (0.071 mL, 0. 88 minol) and 2, 2 ,2-trichloro- 1, 1-diinethylethyl WO 98/50399 WO 9850399PCTIUS98/09385 28 chioroformate (195 mg, 0.80 mmol) in CH,C1 2 followed by diphenyl chiorophosphate (0.213 mL, 1.10 mmol), triethylamine (0.20 mL, 1.46 mmol) and catalytic 4pyrrolidinopyridine (50 mg) to afford 1.10 g (89 of 3-ezlx-S--()3 undecanoyloxytetradecanoylamrno]propyl 2-deoxy-4-O-diphenylphosphono30[(R)-3.

undecanoyloxytetradecanoyl]-6-O-(2,2,2-tricbioro. ,1 -dimethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylamino)-f3-D-glucopyranoside as a colorless amorphous solid: 'H NMR (CDC1 3 860.87 12 H, J= 6.7 Hz), 1. 1 1.6 (in, 72 1.78 3 1.86 (s, 3 2.01 (in, 1 2.18 (mn, 3 2.40 (in, 2 2.67 (in, I 2.88 1 H, J 6.7 Hz), 2.97 1 H, J =6.9 Hz), 3.41 2H), 3.72 1 3.82 1 4.24 1 H), 4.42 1 H, J 11.8 Hz), 4.64 (mn, 3 5.16 (in, I 5.3 9 (in, 2 5.75 I H, J= 4.6 Hz), 6.05 1 H, J 8.4 Hz), 7.22 (mn, 15 H).

In the same manner as described in Example the compound prepared in above (1.0 g, 0.59 iniol) was deprotected with zinc (2.0 g, 30 mniol) and acylated with (R)-3-undecanoyloxytetradecanoic acid (292 mng, 0.71 inmol) in the presence of EEDQ (210 mg, 0.85 minol) to afford 388 mg (40 of 3-benzyloxy-(S)-2- -undecanoyloxytetradecanoylainino~propyI 2 -deoxy-4-O-diphenylphosphono-2- 3 -undecanoyloxytetradecanoylamino].3...o[(R)-3 -undecanoyltetradecanoyl]-p-Dglucopyranoside as a colorless amorphous solid.

In the same manner as described in Example the compound prepared in above (388 mng, 0.24 minol) was hydrogenated in the presence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) and platinum oxide (300mig) in EtOH AcOH (10:1) to afford 65 mg (17 of 3-hydroxy-(S)-2-[(R)-3undecanoyloxytetradecanoylIamino] propylI 2-deoxy-4-O-phosphono-2- undecanoyloxytetradecanoylamino]-.3 -undecanoyloxytetradecanoyll-3..D glucopyranoside triethylanmmonium salt as a white powder: mp l83-184o C; IR (film.) 3306, 2956, 2922, 2852, 1732, 1644, 1550,1467,1377,1164,1106, 1052, 721 cmw'; 'H1 NMR (CDC1 3

CD

3 OD) 6 0. 88 18 H, J 6.8 Hz), 1. 1 1. 7 (in, 117 2.2 -2.7 (in, 12 3.07 6 H, J 7.1 Hz), 3.3 3.9 (in, 13 4.23 (in, I 4.45 1 H, J 8.2 Hz), 5.0 5.3 (in, 4 1 3 C NMR (CDCl 3 6 173.8, 173.5, 173.3, 170.8, 170.5, 170. 1, 10 1.0, 75.5, 73.1, 71.5, 71.3, 70.9, 70.6, 67.8, 61.6, 60.7, 54.4, 5 0.5, 45.8, 41.5, 41.4, 39.5, 34.6, 34.4, 32.0, 31.2, 29.8, 29.7, 29.4, 28.6,26.1, 25.4, 25.1, 22.7, 14.1, 8.6.

WO 98/50399 WO 9850399PCTIUS98/09385 29 Anal. Calcd. for C 9 0

H

174

N

3 01 8 P C, 66.10; H, 10.85; N, 2.57; P, 1.89.

Found: C, 66.34; H, 10.69; N, 2.32; P, 1.99.

EXAMPLE 6 Preparation of 3 -Hydroxy-(S 2 3 -decanoyloxytetradecanoylamninojpropyl 2-Deoxy- 4 -O0- pho s phono -2 -3 -decan o yo xytetrad ecan oy am ino] 3 -0 -3 decanoyloxytetradecanoy]-p3-Dglucopyranoside Triethylammonium Salt (Compound

R,=R

2

=R

3 n-C 9

H,

9 C0, X=Y=O, n=m=q=O, R 4 =R5=R 7 =Rq9=H, R6=OH, p1I

R

8 =P0 3

H

2 In the same manner as described in Example the compound prepared in Example (2.0 g, 3.12 mmol) was acylated with decanoyloxytetradecanoic acid (1.36 g, 3.42 mmol) in the presence of EDC-MeI (1.12 g, 3.74 mmol) and 4-pyrrolidinopyridine (100 mg) in CH 2

CI

2 (40 mL) to afford 2.49 g (79 of 3 -benzyloxy-(S)-2-(allyloxycarbonylamino)propyl 2-deoxy-4,6-Ois o pr op y Iid en e -3 (R)3d ec an o yo x ytetr ad e canoy I] 2 2 trichloroethoxycarbonylamino)--D-glucopyranoside as an amorphous solid: 'H NMR (CDCl 3 )5~0.88 6 H, J= 6.7 Hz), 1. 1 1.6 (in, 34 1.36 3 1.46 3 2.27 2 H, J= 6.9 Hz), 2.48 (dd, 1 H, J= 15.1, 6.0 Hz), 2.60 (dd, 1 H, J 15.1, 6.7 Hz), 3.25 (in, I 3.35 4.0 (in, 9 4.23 (in, I 4.42 (in, 1 4.52 (in, 4 4.95 (in, 2 H), 5.17 3H), 5.88 1 7.36 5 H).

In the same manner as described in Example the compound prepared in above (2.47 g, 2.42 mmol) was deprotected in THF (40 mL) in the presence of diinethyl malonate (2.0 mL, 1.75 minol) and tetrakis(triphenylphosphine)palladium(O) (400 mng) and then acylated with (R)-3-decanoyloxytetradecanoic acid (1.06 g, 2.66 inmol) in the presence of EEDQ (740 mng, 3 minol) to afford 1.86 g (60 of 3benzyloxy-(S)-2-[(R)-3 -decanoyloxytetradecanoylamino]propyl 2-deoxy-4,6-Osopropylidene-3 -decanoy loxytetradecanoyl ,2 ,2trichloroethoxycarbonylamino)-o-D-glucopyranoside as a colorless amorphous solid: 'H NMR (CDCI 3 )8~0.87 12 H, J 6.7 Hz), 1. 1 1.7 (in, 68 1.37 3 1.46 3 2.32 (mn, 4 2.50 (dd, 1 H, J= 15.1, 6.0 Hz), 2.62 (dd, 1 H, J= 15.1, 6.8 Hz), 3.29 (in, 2 3.44 (in, 1 3.55 (in, 1 3.74 (in, 3 3.93 (in, I 4.18 (in, 1 4.34 WO 98/50399 WO 9850399PCTIUJS98/09385 In1 4.57 I H, J1= 11.8 Hz), 4.65 (in, 2 5.01 (in, 2 6.04 1 H, Jl= 8.3 Hz), 7.36 (in, 5 H).

In the same manner as described in Example the compound prepared in above (900 mg, 0.72 minol) was deprotected in 90 aqueous AcOH (40 mL), and then treated with pyridine (0.07 1 mL, 0.88 mmol) and 2 2 2 -trichloro-,l -diinethylethyl chioroformate (195 mng, 0. 80 inmol) in CH 2

CI

2 followed by diphenyl chiorophosphate (0.23 mL, 1.10 minol), triethylamine (0.20 mL, 1.46 minol) and catalytic 4pyrrolidinopyridine (50 mg) to afford 1.05 g (86 of 3-benzyloxy-(S)-2-[(R)-3decanoyloxytetradecanoylamino]propyl 2-deoxy-4-O-diphenylphosphono3..0[(R)3 decanoyloxytetradecanoy)-6-0-(2,2,2-.trichloro 1,1 -dimethylethoxycarbonyl)-2-(2,2,2.

trichloroethoxycarbonylamino)-3D-glucopyranoside as a colorless amorphous solid: 'H NMR (CDCl 3 6 0.87 12 J= 6.3 Hz), 1.1 1.6 68 1.78 3 1.86 (s, 3 2.01 (mn, 1 2.18 (mn, 3 2.40 (mn, 2 2.67 (in, 1 2.88 I H, J Hz), 2.97 1 H, J= 6.9 Hz), 3.41 (mn,2 3.72 I 3.82 1 4.24 1 H), 4.42 1 H, J= 11.8 Hz), 4.64 (in, 3 5.16 (in, 1 5.39 (mn, 2 5.75 1 H, J 4.3 Hz), 6.05 1 H, J 8.4 Hz), 7.22 (mn, 15 H).

In the same manner as described in Example the compound prepared in above (1.0 g, 0.60 inmol) was deprotected with zinc (2.0 g, 30 minol) and acylated with (R)-3-decanoyloxytetradecanoic acid (285 mg, 0.72 inmol) in the presence of EEDQ (210 mg, 0.86 minol) to afford 332 mng (34 of 3-benzyloxy-(S)-2-[(R)-3decanoyloxytetradecanoylaminolpropyl 2-deoxy-4-0-diphenylphosphono-2-[(R)3decanoyloxytetradecanoylanino]-3-)- -decanoyltetradecanoyl] -f-Dglucopyranoside as a colorless amorphous solid.

In the same manner as described in Example the compound prepared in above (332 ing, 0.20 inmol) was hydrogenated in the presence of palladium hydroxide (150 mg) on carbon in EtOH (10 inL) and platinum oxide (3 00 mg) in EtOH AcOH (10:1) to afford 173 mng (55 of 3-hydroxy-(5)-2-[(R)-3decanoyloxytetradecanoylainino]propyl 2 -deoxy-4-0-phosphono-2[(R)-3decanoy loxytetradecanoyl amino] 3 3 -decanoyloxytetradecanoy]

P-D

glucopyranoside triethylammonium salt as a white powder: mp 179-181P C; IR (film) 3295, 2956, 2923, 2853, 1732, 1650, 1555, 1467, 1377, 1320, 1169, 1134, 1104, 1051, WO 98/50399 WO 9850399PCT[US98/09385 31 979, 801, 721 'H NMR(CDC 3

-CD

3 OD) 60.88 18 H,J= 6.9 Hz), 1.1 1.7(in, I1I1- 2.2 2.7 (mn, 12 3.07 6 H, J =6.5 Hz), 3 .3 4.3 (in, 14 4.45 1 H, J 8.0 Hz), 5.0 5.3 (mn, 4 7.39 (in, 1 7.53 1 H, J 9.1 Hz); 1 3 C NMR (CDC1 3 6 173.7, 173.4, 173.2, 170.7, 170.5, 170.1, 101.0, 75.4, 73.1, 71.6, 71.1, 70.8, 70.5,67.8,61.4,60.8, 54.3, 50.4,45.8,41.3-,39.5,34.5,31.9,29.8,29.7,29.4, 25.4,25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for C 87

H,

6 8

N

3 0, 8 P -H 2 0: C, 65.58; H, 10.75; N, 2.64; P, 1.94.

Found: C, 65.49; H, 10.75; N, 2.64; P, 1.97.

EXAMPLE 7 (B6) Preparation of 3-Hydroxy-(S)-2-[(R)-3 -decanoyloxytetradecanoylaminolpropyl 2-Deoxy- 6 pho sphono-2 -3 -dec anoylIo xytetradec an oyaino I -3 decanoyloxytetradecanoyl]-3-D-glucopyranoside Triethylammonium Salt (Compound of R,=R 2

=R

3 n-C 9 H,1 9 C0, X=Y=O, n=in=q=0, R,=R 5

=R

7

=R

8

R

6 =OH, p=l, R9=pO 3

H

2 In the same manner as described in Example the compound prepared in Example (900 ing, 0.72 minol) was deprotected in 90 aqueous AcOH (20 mL).

The residue was dissolved in CH 2 Cl, (20 mL), cooled to 0 0 C, and treated with triethylamnine, 14 mL, 1.0 iniol) and diphenyl chiorophosphate 17 mL, 0.8 inmol).

The mixture was stirred for an additional 6 h, and then quenched with 50 mL of 10 HCl. The product was extracted with EtOAc (3 x 50 mL) and dried over Na 2

SO

4 Chromatography on silica gel with 50 EtOAc hexanes afforded 636 mng (63 of 3benzyloxy-(S)-2- -decanoyloxytetradecanoylainino]propyl 2-deoxy-6-Odiphenylphosphono-3 -decanoyloxytetradecanoyl] trichloroethoxycarbonylamino)-p3-D-glucopyranoside as a colorless amorphous solid: 'H NMR (CDCl 3 60.87 12 H,J= 6.0 Hz), 1.1 1.6 68 1.79 3 1.86 3 2.01 (in, 1 2.18 3H), 2.40 2H), 2.67 1 2.89 I H, J= 6.5 Hz), 2.97 I H, J= 6.9 Hz), 3.41 (in, 2 3.75 (in, 1 3.82 (in, 1 4.24 (in, 1 4.42 1 H, J 11.8 Hz), 4.65 (in, 3 5.16 (in, 1 5.39 (in, 2 5.75 1 H, J 4.3 Hz), 6.05 1 H, J =8.4 Hz), 7.22 (in, 15 H).

In the same manner as described in Example the compound prepared in above (620 g, 0.44 iniol) was deprotected with zinc (722 ing, 11 minol) 11 WO 98/50399 WO 9850399PCTIUS98/09385 32 and acylated with (R)-3-decanoyloxytetradecanoic acid (190 mg, 0.48 mmol) in the presence of EEDQ (170 mg, 0.58 mmol) to afford 254 mg (36 of 3-benzyloxy-(Sy2- 3 -decanoyloxytetradecanoylamnino]propyl 2 -deoxy-6-O-diphenylphosphono-2-[(R)- 3 -decanoyl oxytetradecanoylamninol -3 -decanoyltetradecanoyl] q.Dglucopyranoside as a colorless amorphous solid.

In the same manner as described in Example the compound prepared in above (254 mg, 0. 16 mmol) was hydrogenated in the presence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) and platinum oxide (300 mg) in EtOH AcOH (10:1) to afford 34 mg (13 of 3-hydroxy-(S)-2+[R)-3decanoyloxytetradecanoylamino]propyl 2-deoxy-6-O-phosphono2-[(R).3 decanoyloxytetradecanoylamino]-3 3 -decanoyloxytetradecanoy1]-P.D glucopyranoside triethylammonium salt as a white powder: mp 169 171 0 C; IR (film) 3306,2922,2853, 1732,1644,1548,1467,13-77,1316, 1165,1106, 1 0 5 3 8 56,722cm-1 'H NMR (CDC 3

CD

3 OD) 560.88 18,H, J 6.7 Hz), 1. 1 1.7 (in, Ill 2.2 2.7 (in, 12 3.05 (mn, 6 3.3 -3.95 (in, 12 4.11 (in, 1 4.34 (in, I 4.89 (in, 1 5.0 -5.3 (in, 4 "CNMR (CDC1 3 6 173.8,173.4, 171.1,170.5, 101.3, 75.3, 74.9, 71.2, 71.0,70.6, 68.8,67.3, 65.1, 61.4,53.4,50.7,45.9, 41.5,41.3,39.6,34.6,32.0,29.8, 29.6, 29.4, 25.3, 25.1, 22.7, 14.1, 8.7.

Anal. Calcd. for C 87

HI

68

N

3 01 8 P -H 2 0: C, 65.58; H, 10.75; N, 2.64; P, 1.94.

Found: C, 65.60; H, 10.34; N, 2.36; P, 2.01..

EXAMPLE 8 (B37) Preparation of 3 -Hydroxy-(S)- 2 -[(R)-3-nonanoyloxytetradecanoylan-uno]propyl 2-Deoxy- 4-O-phosphono-2-[(R)-3 -nonanoyloxytetradecanoylaminop.3 nonanoyloxytetradecanoy1]-p-D.glucopyranoside Triethylaminonium Salt (Compound

R,=R

2

=R

3 n-C 8

H,

7 C0, X=YO,

R,

4

=R

5

=R

7

=R

9

R

6 pl,

R

8 =P0 3

H

2 In the same manner as described in Examnple the compound prepared in Example (1.0 g, 1.56 inmol) was acylated with nonanoyloxytetradecanoic acid (660 mng, 1.71 iniol) in the presence of EDC-MeI (560 mg, 1.87 nunol) and 4-pyrrolidinopyridine (50 mg) in CH 2 C1 2 (20 inL) to afford 1.31 g (83 of 3 -benzyloxy-(S)- 2 -(allyloxycarbonylamino)propyI 2-deoxy-4,6-0- WO 98/50399 WO 9850399PCTIUS98/09385 33 i so prop ylIi dene- 3 0- 3 -no nano ylIoxytetr ad ecanoyl1] -2 ,2,2 trichaloroethoxycarbonylamino)-p3-D-glucopyranoside as an amorphous solid: 'H NMR (CDCl 3 5 0. 87 6 H, J 6.8 Hz), 1. 1 1.6 (in, 32 1.3 7 3 1.46 3 2.27 2 H, J= 7.4 Hz), 2.50 (dd, 1 H, 15.1, 6.0 Hz), 2.63 (dd, I H, J= 15.1, 6.8 Hz), 3.26 (i,1 3.35 4.0 9H), 4.32 1 H, J =7.8 Hz), 4.41 1 H, J= 12.0OHz), 4.51 (in, 4 4.95 (in, 2 5.18 (in, 2 5.29 1 H, J =17.2 Hz), 5.88 (in, I 7.36 (mn,

H).

In the same manner as described in Example the compound prepared in above (1.29 g, 1.28 iniol) was deprotected in THF (20 inL) in the presence of diinethyl malonate (1.0 mL, 0.88 inmol) and tetrakis(triphenylphosphine)palladium(0) (200 mg) and then acylated with 3 -nonanoyloxytetradecanoic acid (540 mg, 1.41 inmol) in the presence of EEDQ (370 ing, 1.5 minol) to afford 1.02 g (65 of 3benzyloxy-(S)-2 -3 -nonanoy loxytetradecanoy Iamnino] propy I 2-deoxy-4,6-0isopropyl idene-3 -nonanoyloxytetradecanoyl] -2 trichloroethoxycarbonylamino)-V.Dglucopyranoside as a colorless amorphous solid: 1H NMR (CDC1 3 )80.87 12 H, J= 6.1 Hz), 1. 1- 1.7 (in, 64 1.37 3 1.46 (s, 3 2.28 (in, 4 2.50 (dd, 1 H, J 15.5, 6.0 Hz), 2.62 (dd, 1 H, J 14.8, 6.3 Hz), 3.27 (in, 2 3.44 (in, I 3.55 (in, 1 3.74 (in, 3 3.93 (in, 1 4.18 (in, I H), 4.34 (in, 2 4.57 1 H, J= 11.8 Hz), 4.65 (in, 2 4.97 I H, J= 9.6 Hz), 5.06 1 H, J 8.6 Hz), 5.15 (in, 2 6.05 1 H, J =8.2 Hz), 7.3 5 (mn, 5 H).

In the same manner as described in Example the compound prepared in above 0 g, 0. 81 inmol) was deprotected in 90 aqueous AcOH (20 mnL), treated with pyridine (0.080 mL, 0.98 mmol) and 2, 2 ,2-trichloro-1,1-dimethylethyI chioroforinate (215 ing, 0.89 inmol) in CH 2 Cl 2 followed by diphenyl chiorophosphate (0.25 inL, 1.22 inmol), triethylamnine (0.21 inL, 1.52 iniol) and catalytic 4pyrrolidinopyridine (50 mg) to afford 1.17 g (87 of 3-benzyloxy-(S)-2-[R)-3nonanoyloxytetradecanoylamino]propyl 2 -deoxy- 4 -O-diphenylphosphono-3 nonanoyloxytetradecanoyl]p6-0(2,2,2trichloro.. ,1 -diinethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylanino)-p3-D-glucopyranoside as a colorless amorphous solid: 1H NMR (CDC1 3 )8 0.87 12 H, J =6.1 Hz), 1. 1- 1. 6(m, 64 1.78 3 l. 86 (s, 3 2.01 (in, I 2.18 (in, 3 2.40 (mn, 2 2.67 (in, 1 2.88 1 H, J WO 98/50399 WO 9850399PCTAUS98/09385 34 Hz), 2.97 1 H, J= 6.9 Hz), 3.41 (in, 2 3.72 (in, I 3.82 (in, 1 4.24 (in, 1 H), 4.42 I H, J 11. 8 Hz), 4.64 (in, 3 5.16 (in, I 5.3 9 (in, 2 5.75 1 H, J= 4.3 Hz), 6.05 1 H, J 8.4 Hz), 7.22 (in, 15 H).

In the same manner as described in Example the compound prepared in above (1.1I g, 0.66 inmol) was deprotected with zinc (2.2 g, 33 inmol) and acylated with 3 -nonanoyloxytetradecanoic acid (305 mg, 0.79 minol) in the presence of EEDQ (235 mg, 0.95 minol) to afford 373 mng (35 of 3-benzyloxy-(S)-2+[R)-3 nonanoyloxytetradecanoylamino]propyI 2-deoxy-4-O-diphenylphosphono-2-[(R)3 nonanoyloxytetradecanoylamino]-3..o..[(R)-3 -nonanoyltetradecanoylL.WD.

glucopyrano side as a colorless amorphous solid.

In the same manner as described in Example the compound prepared in above (373 mng, 0.23 mmol) was hydrogenated in the presence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) and platinum oxide (300 mng) in EtOH AcOH (10:1) to afford 43 mng (12 of 3-hydroxy-(S)-2+[R)-3nonano ylIoxytetradecanoylIamino] propylI 2-deoxy-4-O-phosphono-2.[(R).3 nonanoyloxytetradecanoyl amino] -3 3 -no nanoy Ioxytetradecanoyl] 0 D glucopyranoside triethylarmonium. salt as a white powder: mp 176-179o C; IR (film) 3298, 2956, 2923, 2853, 1733, 1646, 1551, 1467,1337,1316, 1254, 1166, 1106, 1053, 722 'H NMR (CDC1 3

CD

3 OD) 8 0.87 18 H, J 6.7 Hz), 1. 1 1.7 (in, 105 H), 2.2 2.7 (in, 12 3.03 6 H, J 7.0 Hz), 3.3 -4.3 (in, 14 4.43 I H, J 7.1 Hz), 5.0 -5.3 (in, 4 7.12 1 H, J 7.7 Hz), 7.17 1 H, J= 8.2 Hz); 3 C NMR (CDCl 3 8 173.9, 173.5, 173.3, 170.8, 170.5, 170.1, 100.9, 75.5, 73.1, 71.4, 71.1, 70.9, 70.6, 67.8, 61.6, 60.7, 54.3, 50.5, 45.8, 41.6, 41.4, 39.5, 34.6, 34.4, 32.0, 31.9, 29.8, 29.4, 29.3, 25.4, 2 5.1, 22.7, 14.1, 8.6.

Anal. Caled. for C 8 8

H,

64

N

3 0, 8 p: C, 65.8 1; H, 10.65; N, 2.74; P, 2.02. Found: C, 66.14; H, 10-46; N, 2.58; P, 1.84.

WO 98/50399 WO 9850399PCT/US98/09385 EXAMPLE 9 (B8) Proparation of 3 -Hydroxy-(S)-2-[(R)-3 -heptanoyloxytetradecanoylamino]propyl 2- D eoxy-4-0O-phosphono-2 3 -heptanoylIoxytetradecanoyl amino] 3 heptanoyloxytetradecanoy]-p3-D-glucopyranoside Triethylammonium Salt (Compound

R,=R

2

=R

3 n-C 6 H,1 3 C0, X=Y=O, n=m=q=0, R 4

=R

5

=R

7

=R

9

R

6 =OH, p=1,

R

8 =P0 3

H

2 In the same manner as described in Example the compound prepared in Example (1.0 g, 1.56 mmol) was acylated with heptanoyloxytetradecanoic acid (610 mg, 1. 71 mmol) in the presence of EDC-MeI (560 mg, 1.87 minol) and 4-pyrrolidinopyridine (50 mg) in CH 2

CI

2 (20 mL) to afford 1.24 g (82 of 3 -benzyloxy-(S)-2-(allyloxycarbonylamino)propyI 2-deoxy-4,6-0sopropy li dene -3 0- 3 -heptanoyloxytetradecanoylI] -2 trichloroethoxycarbonylamino)-o3-D-glucopyranoside as an amorphous solid: 'H NMR (CDC1 3 8 0.88 6 H, J= 6.0 Hz), 1. 1 1.6 (in, 28 1.38 3 1.47 3 2.29 2H, J=7.4 Hz), 2.51 (dd, I H, J= 15.1, 6.0OHz), 2.63 (dd, I H, J= 15.1, 6.8 Hz), 3.26 (i,1 3.35 4.0 (in, 9 4.32 1 H, J 7.3 Hz), 4.41 1 H, J= 12.0 Hz), 4.51 (i,4 4.95 (mn, 2 5.18 (in, 2 5.29 1 H, J= 17.3 Hz), 5.88 (in, I 7.36 (in,

H).

In the same manner as described in Example the compound prepared in above (1.22 g, 1.25 mmol) was deprotected in THF (20 inL) in the presence of diinethyl malonate (1.0 mL, 0.88 inmol) and tetrakis(triphenylphosphine)palladiuin(0) (200 ing) and then acylated with (R)-3-heptanoyloxytetradecanoic acid (490 ing, 1.38 iniol) in the presence of EEDQ (370 mg, 1.5 mmol) to afford 925 mg (62 of 3benzyloxy-(S)-2- -heptanoyloxytetradecanoylaminolpropyl 2-deoxy-4 ,6-Oisopropylidene-3-0-[(R)-3-heptanoyloxytetradecanoylj.2-(2,22trichloroethoxycarbonylamino)-3-D-glucopyranoside as a colorless amorphous solid: 'H NMR (CDC1 3 860.87 12 H, J= 6.7 Hz), 1. 1 1.7 (in, 56 1.37 3 1.46 3 2.32 (in, 4 2.50 (dd, 1 H, J =15.1, 6.0 Hz), 2.62 (dd, 1 H, J= 15.1, 6.8 Hz), 3.29 (in, 2 3.44 (in, 1 3.55 (in, 1 3.74 (in, 3 3.93 (in, 1 4.18 (in, I 4.34 (in, 1 4.5 7 1 H, J =11. 8Hz), 4.65 (in, 2 5.01 (in, 2 6.04 1 H, J =8.3 Hz), 7.36 5 H).

WO 98/50399 WO 9850399PCT/US98/09385 36 In the same manner as described in Example the compound prepared in above (920 mg, 0.76 mmol) was deprotected in 90 aqueous AcOH (20 mL), and then treated with pyridine (0.075 mL, 0.92 mmol) and 2 2 2 -trichloro-1,lI-dimethylethyl chloroformate (200 mg, 0.84 mmol) in CH 2 Cl, followed by diphenyl chlorophosphate(0.24 mL, 1. 14 mmol), triethylamine (0.21 mL, 1.52 mmol) and catalytic 4-pyrrolidinopyridine (50 mg) to afford 1.03 g (83 of 3-benzyloxy-(S)-2-[(R)-3heptanoyloxytetradecanoylamino]propyl 2-deoxy-4-O-diphenylphosphono.3..0-[(R).3 heptanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro- 1, 1 -dimethylethoxycarbonyl)-2-(22,2 trichloroethoxycarbonylanino)-13-D-glucopyranoside as a colorless amorphous solid: 'H NMR (CDCl 3 80.87 12 H, J= 6.3 Hz), 1. 1- 1.6 56 1.78 3 1.86 (s, 3 2.01 (in, I 2.18 (mn, 3 2.40 (in, 2 2.67 (mn, 1 2.88 1 H, J= Hz), 2.97 1 H, J= 6.9 Hz), 3.41 2H), 3.72 1 3.82 I 4.24 1 H), 4.42 I H, J= 11.8 Hz), 4.64 (in, 3 5.16 (in, 1 5.39 (mn, 2 5.75 1 H, J= 4.3 Hz), 6.05 1 H, J= 8.4 Hz), 7.22 (mn, 15 H).

In the same manner as described in Example the compound prepared in above (1.0 g, 0.61 inmol) was deprotected with zinc (2.0 g, 31 mmol) and acylated with -heptanoyloxytetradecanoic acid (260 mg, 0.73 inmol) in the presence of EEDQ (220 mg, 0.88 imol) to afford 203 mg (21 of 3-benzyloxy-(S)-2-[(R)-3heptanoyloxytetradecanoylainino]propyl 2-deoxy-4-O-diphenylphosphono-2-[(R)-3 heptanoyloxytetradecanoylamino] -3 3 -heptanoyloxytetradecanoyl] -j-Dglucopyranoside as a colorless amorphous solid.

In the same manner as described in Example the compound prepared in above (203 mng, 0. 13 minol) was hydrogenated in the presence of palladium hydroxide (100 mg) on carbon in EtOH (10 mL) and platinum oxide (200 mg) in EtOH AcOH (10:1) to afford 39 mng (21 of 3-hydroxy-(S)-2-[(R)-3heptanoyloxytetradecanoylamino]propyl 2-deoxy-4-O-phosphono-2-[(R)-3 heptanoyloxytetradecanoylamino] 3 -O-[(R)-3-heptanoyloxytetradecanoyl] -1-Dglucopyranoside triethylammonium salt as a white powder: mp 171-172' C; IR (film) 3305, 2955, 2924, 2853, 1734, 1644, 1553, 1466,1377,1170,1102, 1052, 722 cm- 1

'H

NMR (CDC 3

CD

3 OD) 8 0.88 (in, 18 1. 1 1.7 (in, 93 2.2 2.7 (in, 12 3.06 6 H, J 7.1 Hz), 3.3 4.0 (in, 13 4.23 1 H, J 9.3 Hz), 4.43 1 H, J= 8.2 I. WO 98/50399 PCTIUS98/09385 37 Hz), 5.0 -5.3 (in, 4 7.30 1 H, J= 8.5 Hz), 7.43 I H, J =8.5 Hz); B 3 C NMR (CDC1 3 8 173.8, 173.5, 173.2, 170.8, 170.5, 170.2, 101.0, 77.2, 75.5, 73.1, 71.6, 7 1.1, 70.9, 70.6, 67.8,61.6,60.8, 54.4, 50.5,45.8,41.6,41.4, 39.5, 34.6, 34.4,32.0, 31.6, 29.8, 29.6, 29.4, 28.9, 25.4, 25.1, 22.7, 22.6, 14.1, 8.6.

Anal. Caled. for C 7 8

H,

5

ON

3 0, 8 P C, 63.86; H, 10.44; N, 2.86; P, 2.11.

Found: C, 63.47; H, 10.20; N, 2.59; P, 2.02.

EXAMPLE 10 (B39) Preparation of 4-Hydroxy.-(S)-3 -decanoyloxytetradecanoyl]butyl 2-Deoxy-4-Opho sphono 2 3 -decanoyo xytetradecanoy lanino]3 -3 decanoyltetradecanoyl-p3-D-glucopyranoside Triethylamnmonium Salt (Compound

R,=R

2

=R

3 n-C 9

H,

9 C0, X=Y=O, n=p=1, m=q=0, R, 4

=R

5 =R7=R 9

R.

6 =01H, R 8 =P0 3

H

2 In the same manner as described in Example the compound prepared in Example (3.1 g, 5.9 mmol) and (R)-3-(allyloxycarbonylamino)-4benzyloxy- 1 -butanol (1.1I g, 3.94 inmol) were coupled in the presence of boron trifluoride etherate (3.0 mL, 23.6 mmol) to afford 1.96 g (67 of 4-benzyloxy-(S)-3- (allyloxycarbonylamino)butyl 2-deoxy-3 ,4,6-tri-O-acetyl-2-(2,2,2trichloroethoxycarbonylamino)-p-D-glucopyranoside as an amorphous solid. In the same manner as described in Example the compound prepared above (1.8 g, 2.43 mmol) was deacylated in methanol (25 mL) with ammonium hydroxide (5 mL) and then treated with 2,2-dimethoxypropane (25 mL) and camphorsulfonic acid (100 mg) to afford 1.34 g (84 of 4 -benzyloxy-(S)-3-(allyloxycarbonylamino)butyl 2-deoxy-4,6-Oisopropylidine- 2 2 ,2,2-trichloroethoxycarbonylamino)-.-D-glucopyranoside.

In the same manner as described in Example the compound prepared in above (1.0 g, 1.53 inmol) was acylated with decanoyloxytetradecanoic acid (670 mg, 1.68 mmol) in the presence of EDC'MeI (550 mng, 1.85 mn-ol) and 4-pyrrolidinopyridine (50 mng) in CH 2 Cl 2 (15 mL) to afford 1.03 g (65 of 4 -benzyloxy-(S)-3-(allyloxycarbonylamino)butyl 2-deoxy-4,6-Oi is opro pyIi dene -3 -0O4[(R-3-d e can oy loxy tetradec anoy I -2 -(2,2,2 30 trichloroethoxycarbonylamino)-3-D-glucopyranoside as an amorphous solid: 'H NMR (CDCl 3 860.88 6 H, J= 6.9 Hz), 1.1 1.6 (in, 34 1.37 3 1.47 3 1.85 (in, 2 2.28 2 H,J 7.6 Hz), 2.50 (dd, 1 H, J 15.1, 6.0 Hz), 2.63 (dd, 1 H,J WO 98/50399 WO 9850399PCTIUS98/09385 38 15.1, 6.7 Hz), 3.30 (in, 1 3.49 (mn, 4 3.68 1 H, J1=9.4 Hz), 3.77 I H, J= 10.4 Hz), 3.92 (in, 3 4.54 (in, 5 4.69 (in, 2 5.1 5.4 (mn, 4 5.91 (in, 1 H), 7.33 (in, 5 H).

In the same manner as described in Example the compound prepared in above (1.0 g, 0.97 inmol) was deprotected in THF (20 mL) in the presence of diinethyl malonate (1.0 mL, 0.88 minol) and tetrakis(triphenylphosphine)palladium(0) (200 ing) and then acylated with (R)-3-decanoyloxytetradecanoic acid (425 ing, 1.07 mmol) in the presence of EEDQ (317 mg, 1.28 inmol) to afford 660 mng (51 of 4benzylIoxy 3 -3 decanoy loxytetradecanoyl amino] propyl 2-deoxy-4,6-Oisopropylidene-3-O-[(R)-3-decanoyloxytetradecanoyl]-2-.(2,2,2..

trichloroethoxycarbonylamlino)-3-D-glucopyranoside as a colorless amorphous solid: 'H NMR (CDCl 3 6 0.88 12 H, J= 6.6 Hz), 1. 1 1.7 (in, 68 1.37 3 1.47 3 2.26 2 H, J 7.1 Hz), 2.41 (in, 2 2.62 (dd, I H, J =14.9, 6.4 Hz), 3.29 (in, 1 3.48 (in, 3 3.71 (in, 2 3.92 (in, 2 4.18 (in, 1 4.49 (in, 2 4.68 2 H, J= 11.5 Hz), 5.15 (in, 2 5.55 I H, J= 8.8 Hz), 6.17 1 H, J= 7.2 Hz), 7.32 (in, 5 H).

In the same manner as described in Example the compound prepared in above (640 ing, 0.48 inmol) was deprotected in 90 aqueous AcOH (20 mL), and then treated with pyridine (0.047 inL, 0.58 inmol) and 2,2,2-trichloro-1, I-diinethylethyl chioroforinate (127 mg, 0.53 minol) in CH,C1 2 followed by diphenyl chiorophosphate 15 mL, 0.72 minol), triethylamine 13 mL, 0.96 inmol) and catalytic 4pyrrolidinopyridine (50 ing) to afford 389 mng (47 of 4-benzyloxy-(S)-3-[(R)-3decanoyloxytetradecanoyllbutyl 2-deoxy-4-O-diphenylphosphono-3 decanoyloxytetradecanoyl]-6-O..(2,2,2.trichloro- 1,1-diinethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylamino)3..D.glucopyranoside as a colorless amorphous solid: NMR (CDCl 3 6 0.88 12 H, J= 6.6 Hz), 1. 1 1.6 (in, 68 1.79 3 1.86 (s, 3 2.22 (in, 4 2.40 (in, 4 3.49 (mn, 4 3.78 (in, I 3.93 (in, 1 4.1 (in, 5 4.9 -4.6 (in, 4 5.13 (in, 2 5.5 1 I H, J 8.9 Hz), 5.84 1 H, J= 6.9 Hz), 6.09 I H, J 8.0 Hz), 7.26 (in, 15 H).

In the same manner as described in Example the compound prepared in above (375 g, 0.23 inmol) was deprotected with zinc (752 mng, 11.5 WO 98/50399 WO 9850399PCT/US98/09385 39 mmol) and acylated with 3 -decanoyloxytetradecanoic acid (10 1 mg, 0.25 mmol) in the presence of EEDQ (70 mg, 0.28 mmol) to afford 270 mg (67 of 4-benzyloxy-(S)- -decanoyloxytetradecanoyljbutyl 2-deoxy-4-O-diphenylphosphono-2-.[(R)-3 -decanoyltetradecanoyl]-3..D.

glucopyranoside as a colorless amorphous solid.

In the same manner as described in Example the compound prepared in above (270 mg, 0.15 mmol) was hydrogenated in the presence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) and platinum oxide (300 mg) in EtOH AcOH (10:1) to afford 93 mg (39 of 4-hydroxy-(S)-3-[(R)-3decanoyloxytetradecanoyllbutyl 2-deoxy-4-O-phosphono-2-[(R)-3.

decanoyloxytetradecanoylamino~j.3 3-decanoyltetradecanoyl] -f-Dglucopyranoside triethylammonium salt as a white powder: mp 179-1811 C (dec): IR (film) 3287, 2956, 2923, 2853, 1734, 1654, 1 552, 1466, 1378, 1246, 1164, 1106, 1085, 1052,721 'H NMR (CDC1 3

-CD

3 OD) 60.88 18 H, J=6.9 Hz), 1. 1- 1.7 I11 2.2 2.7 (in, 14 3.06 6 H, J= 6.9 Hz), 3.2 4.0 (in, 13 4.21 (mn, 1 4.50 1 H,.J=7.7 Hz), 5.0 -5.3 4H), 7.11 2H); 1 3 CNMR (CDC1 3 8 173.8, 173.5, 173.3, 170.9, 170.5, 170.1, 101.1, 77.2, 75.5, 72.8, 71.3, 71.0, 70.6, 66.4, 64.0, 60.7, 54.8, 50.2,45.8, 41.6,39.5, 34.6, 34.5, 34.4, 32.0,30.6,29.8,29.7, 29.6,29.5, 29.4, 25.4, 25.1, 22.7, 14.2, 8.6.

Anal. Calcd. for C 8 8 H1 70

N

3 0 18 P: C, 66.65; H, 10.78; N, 2.64; P, 1.95. Found: C, 66.65; H, 10.68; N, 2.50; P, 1.94.

EXAMPLE 11I (B Preparation of 4 -Hydroxy-(S)- 2 -[(R)-3-decanoyloxytetradecanoyl]butyI 2-Deoxy-4-Opho sphono 2 3 -decan oyloxytetrad ecan oylarn ino -3 decanoyltetradecanoyl]-p-D-glucopyranoside Triethylanimoniuin Salt (Compound

R

1

=R

2

=R

3 n-C 9 H 1 19 C0, X=Y=O,

R

4

=R

5

=R

7

=R

9

R

6 =OH, p=2, R 8 =P0 3

H

2 In the same manner as described in Example the compound prepared in Example (5.1 g, 9.7 iniol) and (R)-2-(allyloxycarbonylamino)-4benzyloxy-1I-butanol 8 g, 6.45 inmol) were coupled in the presence of boron trifluoride etherate (4.9 inL, 38.0 inmol) to afford 2.92 g (61 of 4-benzyloxy-(SJ-2- (allylIoxycarb onyl am ino)propylI 2 -deoxy -3,4,6-tri-O-acetyl..2-(2,2,2- WO 98/50399 WO 9850399PCTIUS98/09385 trichloroethoxycabonylamino)$D-glucopyranoside as an amorphous solid. In the same maimer as described in Example the compound prepared above (2.6 g, 3.51 mmol) was deacylated in methanol (3 5 mL) with ammonium hydroxide (7 mL) and then treated with 2 2 -dimethoxypropane (3 5 mL) and camphorsulfonic acid (100 mg) to afford 1.9 g (72 of 4 -benzyloxy-(S)-2-(allyloxycarbonylamino)butyI 2-deoxy-4,6-Oisopropylidine-2-(2,2,2-trichloroethoxycarbonylamino)-3..D-glucopyranoside.

In the same manner as described in Example the compound prepared in above (1.0 g, 1.53 mmol) was acylated with decanoyloxytetradecanoic acid (670 mg, 1.68 mmol) in the presence of EDC-MeI (550 mg, 1.85 mmol) and 4-pyrrolidinopyridine (50 mg) in CH,C 2 (15 mL) to afford 1.28 g (81 of 4 -benzyloxy-(S)-2-(allyloxycarbonylamino)butyI 2-deoxy-4,6-Oi sop rop yIi de ne -3 -3 de can oyl ox.yte trade canoy I -2 trichloroethoxycarbonya.ino)-o-Dglucopyranoside as an amorphous solid: 'H NMR (CDC1 3 0.88 6H, J =6.9 Hz), 1. 1- 1.7 34 1.37 3 1.47 3 1.82 2H), 2.28 2H, J =7.7 Hz),2.50 (dd, 1 H, J= 15.3, 6.0OHz), 2.63 (dd, 1 H,J 15.2, 6.7 Hz), 3.16 (in, I 3.56 (mn, 3 3.65 I H, J1=9.6 Hz), 3.75 1 H, J 10.4 Hz), 3.88 (mn, 4 4.32 1 H, J= 8.5 Hz), 4.46 2 4.54 (mn, 2 4.67 (in, 2 4.90 (in, 1 5.26 (in, 3 5.89 (in, 1 7.33 (in, 5 H).

In the same manner as described in Example the compound prepared in above (1.25 g, 1.21 inmol) was deprotected in THF (20 mL) in the presence of diinethyl inalonate (1.0 inL, 0.88 inmol) and tetrakis(triphenylphosphine)palladium(O) (200 ing) and then acylated. with (R)-3-decanoyloxytetradecanoic acid (530 ing, 1.33 mmol) in the presence of EEDQ (362 mg, 1.46 mmol) to afford 1. 16 g (72 of 4benzyloxy-(S)-3 3 -decanoyloxytetradecanoylaminolpropyI 2-deoxy-4,6-Oisopropylidene-3-O-[(R)-3-decanoyloxytetradecanoylI>(2,2,2.

trichloroethoxycarbonylamino)-3-D-glucopyranoside as a colorless amorphous solid: 1H NMR (CDC1 3 6 0.88 12 H, J= 6.4 Hz), 1.1 1.7 (mn, 68 1.37 3 1.45 (s, 3 2.26 2 H, J= 7.4 Hz), 2.34 (in, 1 2.50 (dd, 1 H, J= 15.1, 6.0 Hz), 2.62 (dd, 1 H, J= 15.4, 6.3 Hz), 3.12 (in, 1 3.5 -3.95 (mn, 7 4.14 (mn, I 4.29 I H, J 8. 0 Hz), 4.67 (in, 2 4.86 1 H, J =9.6 Hz),5.15 (in, 2 6.16 1 H, J 8.3 Hz), 7.35 (in, 5 H).

I- WO 98/50399 WO 9850399PCTIUS98/09385 41 In the same manner as described in Example the compound prepared in above (1.1 g, 0. 83 mmol) was deprotected in 90 aqueous AcOH (20 mL), and then treated with pyridine (0.080 mL, 1.0 mmol) and 2,2,2-trichioro- 1,1 -dimethylethyl chioroformate (220 mg, 0.91 mmol) in CHICl 2 followed by diphenyl chiorophosphate (0.26 mL, 1.25 mmol), triethylamine (0.23 mnL, 1.66 mnmol) and catalytic 4pyrrolidinopyridine (50 mg) to afford 802 mg (56 of 4-benzyloxy-()-2-[(R)-3decanoyloxytetradecanoylbutyl 2 -deoxy-4-O-diphenylphosphono-3 decanoyloxytetradecanoyl]-6-0-(2,2,2-trichloro 1,1 -dimethylethoxycarbonyl)-2-(2,2,2.

trichloroethoxycarbonylamino)-I3-D-glucopyranoside as a colorless amorphous solid: 1H NMR (CDCl 3 60.87 12 H, J= 6.8 Hz), 1.1 -1.6 (in, 68 1.79 3 1.88 (s, 3 2.23 (in, 4 2.37 (mn, 4 3.57 (in, 4 3.83 (in, 1 4.29 (in, 3 4.44 (in, 2 4.69 (in, 4 5.14 (in, 4 5.62 I H, J =7.6 Hz), 6.15 1 H, J 8.3 Hz), 7.25 (in, 15 H).

In the same manner as described in Example the compound prepared in above (750 ing, 0.43 inmol) was deprotected with zinc (1.42 g, 21.7 iniol) and acylated with -decanoyloxytetradecanoic acid (190 mng, 0.48 mmol) in the presence of EEDQ (130 mg, 0.53 inmol) to afford 483 mg (64 of4-benzyloxy-(S)- 2- [(R)-3-decanoyloxytetradecanoyl]butyl 2-deoxy-4-0-diphenylphosphono-2-[(R)-3 decanoyl oxytetradecano y Iamino] -3 0- decanoyltetradecano y] 3-Dglucopyranoside as a colorless amorphous solid.

In the same manner as described in Example the compound prepared in above (483 ing, 0.27 inmol) was hydrogenated in the presence of palladium hydroxide (150 mg) on carbon in EtOH (10 mL) and platinum oxide (300 mng) in EtOH AcOH (10:1) to afford 238 mng (55 of 4-hydroxy-(5)-2-[R)-3decanoyloxytetradecanoyl]butyl 2-deoxy-4-0-phosphono-2-[(R)-3decanoyloxytetradecanoylIamino] 3 3 decanoyItetradecanoyl] 0 Dglucopyranoside triethylammoniuin salt as a white powder: mnp 1 81-1830 C (dec): IR (film) 3294, 2956, 2923, 2853, 1732, 1650, 1556,1466,1377,1320, 1246, 1172, 1108, 1082,1058,859,721 cin'; 'H NMR (CDC 3

CD

3 OD) 5 0.88 18 H,J= 6.9 Hz), 1.1 1. 7 (in, 111 2.2 -2.7 (in, 14 3.06 6 H, J= 7.1 Hz), 3.2 0 (in, 13 4.21 (in, 1 4.46 I H, J= 8.3 Hz), 5.0 -5.3 (mn, 4 1 3 CNMR (CDCl 3 8 173.9, 173.4, I- WO 98/50399 PCT[US98/09385 42 173.2, 171.2, 170.7, 101.0,77.2, 75.4, 73.1,71.4,71.3, 71.1,70.9,70.6, 60.7, 58.4, 54.7, 46.3,45.9,41.6,41.1,39.7,34.8,34.6, 34.4,31.9,29.8,29.6,29.5,29.3,25.4,25.3,25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for C 88

H,

7 0

N

3 0, 8 P: C, 66.5 1; H, 10.78; N, 2.64; P, 1.95. Found: C, 66.8 1; H, 10.68; N, 2.53; P, 1.79.

EXAMPLE 12 (BI 11) Preparation ofN-[(R)-3-Tetradecanoyloxytetradecanoyl]-O-[2Deoxy4-0-phosphono-2- [()3ttaeaolxttaeaol~io-30[R--erdcnyoyerdcny] f-D-glucopyranosyl]-L-serine Triethylammonium Salt (Compound RI=R 2

=R

3 =n-

C,

3 H1 27 C0, X=Y=O, n=m=p=q=0, R 4

=R

5

=R

7

=R

9

R

6

=CO

2 H, R 8 =P0 3

H

2 In the same mannier as described in Example L-serine benzyl ester (0.212 g, 1.08 mmol) was acylated with (R)-3-tetradecanoyloxytetradecanoic acid (0.541 g, 1. 19 mmol) in the presence of EDC&MeI (0.3 53 g, 1. 19 mmol) to give 0.642 g (94%) ofN-(R)-3-tetradecanoyloxytetradecanoyl]-L-serine benzyl ester as a waxy solid: mp 56- 6 1 C; 'H NMR (CDC1 3 6 0.88 6 H, 1= 7Hz), 1. 1- 1.7 (in, 42 2.29 2 H, Hz), 2.50 (in, 2 3.87 (br t, I 3.95 (in, 2 4.65 (in, 1 5.1-5.25 (in, 3 6.69 1 H, J=7 Hz), 7.34 (br s, 5 H).

In the same manner as described in Example the compound prepared in above 19 g, 0.30 inmol) and the compound prepared in Example 2-(4) (0.635 g, 0.478 mmol) were coupled in the presence of mercury cyanide (0.3 g 1.2 mmol) to give 0.425 g ofN-[(R)-3-tetradecanoyloxytetradecanoyl]-O-[2-deoxy-4- O-diphenylphosphono-3 -O-[(R)-3-tetradecanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro- 1, 1 -dimethylethoxy carbonyl)- 2 -tri ch o ro ethoxy carbonyl amino)- P-Dglucopyranosyl]-L-serine benzyl ester as an amorphous solid.

In the same manner as described in Example the compound prepared in above (0.405 g, 0.22 inmol) was deprotected with zinc (0.72 g, I11 mmol) and acylated with (R)-3-tetradecanoyloxytetradecanoic acid (0.12 g, 0.26 mmol) in the presence of EEDQ (0.082 g, 0.33 minol) to give 0.277 g of tetradecanoyloxytetradecanoyl] -0-[2-deoxy-4-O-diphenylphosphono-2- tetradecanoyloxytetradecanoylamino]-3 -0-[(R)-3-tetradecanoyloxytetradecanOyl-p3-Dglucopyranosyl]-L-serine benzyl ester as an amorphous solid: 'H NMR (CDC1 3 8 0.88 WO 98/50399 WO 9850399PCTIUS98/09385 43 18 H, J= 6.5 Hz) 1.0- 1.75 (in, 126 2.15-2.45 (in, 10 2.53 (dd, 1 H, J= 14.7, 4Hz), 2.67 (dd, 1 H, J=14, 6.0 Hz), 3.25 (br t, I H, .k7 Hz), 3.3 5-3.75 (mn, 4 3.88 (dd, 1 H, J= 11.1 Hz), 4.23 dd, 1 H, J= 1.1, 3Hz), 4.6-4.75 (mn, 2 5.03 1 H, J=8.1 Hz), 5.05-5.25 (in, 4 5.48 1 H, J= 10 Hz), 6.40 1 H, J=7.5 Hz), 7.01 1 H, J=8.1 Hz), 7.1-7.4 (in, 15 H).

In the same manner as described in Example the compound prepared in above (0.253 g, 0.133 minol) was hydrogenated in the presence of palladium on carbon (50 mg) and platinum oxide (120 ing) to give 0. 155 g of N- 3 -tetradec ano ylIoxytetradec anoylI] 0- [2 -de oxy -4-0-pho sphono-2 tetradecanoyloxytetradecanoylamino] 3 -O-[(R)-3-tetradecanoyloxytetradecanoylf3.Dglucopyranosyl]-L-serine triethylainmonium salt as a colorless solid: mp 1NOT 0 (dec); IR (film) 3322, 2956, 2924, 2852, 1736, 1732, 1681, 1673, 1667, 1660, 1651, 1467, 1456, 1247, 1174,1110, 1081 11-1NMR (CDCl 3

-CD

3 OD) 80.88 18 H, J= 7 Hz), 1.7 (in, 135 2.2-2.75 (in, 12 3.05 6 H, J=7 Hz), 3.30 (br s, 13 3.7-3.9 (in, 3 3.96 1 H, J=12 Hz), 4.05-4.3 (in, 2 4.34 (in, 1 4.53 1 H, J=7.8 Hz), 5.05-5.3 (mn, 4 7.25-7.35 (in, 2 1 3 C NMR (CDC1 3 8 173.4, 173.2, 171.0, 170.3, 170.2, 169.9, 169.8, 100.8, 75.1,73.4, 71.1, 70.7,70.4, 70.3,60.2, 54.3,45.6,41.2,41.1, 39.2, 34.6, 34.4, 34.2, 32.0, 29.8, 29.5, 25.4, 25.2, 22.7, 14.2, 8.6.

Anal. Calcd for C 99

H-

19 0

N

3 01 9 P -5 H 2 0: C, 64.35; H, 10.91; N, 2.27; P, 1.68.

Found: C, 64.16; H, 10.92; N, 2.37; P, 1.91.

EXAMPLE 13 (B12) Preparation of N-[(R)-3-Dodecanoyloxytetradecanoyl]--[2-deoxy-4O-phosphono2 [(R)-3-dodecanoyloxytetradecanoylaiino]-3..-[(R).3 -dodecanoyloxytetradecanoy]-p3-Dglucopyranosyl]-L-serine Triethylanimonium Salt (Compound R,=R 2

=R

3 =n- C I H 23 C0, X=Y=O,

R

4

=R

5

=R

7

=R

9 R6=CO 2 H, R 8 =P0 3

H

2 In the same manner as described in Example L-serine benzyl ester (390 ing, 2.0 imol) was acylated with (J?)-3-dodecanoyloxytetradecanoic acid (935 mg, 2.2 inmol) in the presence of EDC-MeI (745 mng, 2.5 mmiol) in CH 2 CI, to afford 1.08 g (90 of N-[(J?)-3-dodecanoyloxytetradecanoyl].L-.serine benzyl ester: mp 53-54 TC.

'H NMR (CDCl 3 8 0.88 6 H, J= 6.5 Hz), 1. 1 1.6 (mn, 46 2.30 2 H, J= 7.7 Hz), WO 98/50399 PCT/US98/09385 44 2.50 2 H, 5.6 Hz), 2.62 1 H, J= 6.2 Hz), 3.97 2 4.65 1 5.19 3 6.63 1 H, J= 6.8 Hz), 7.35 (br s, 5 H).

In the same manner as described in Example the compound prepared in Example (1.0 g, 2.02 mmol) was acylated with dodecanoyloxytetradecanoic acid (946 mg, 2.22 mmol) in the presence of EDCMeI (720 mg, 2.4 mmol) and 4-pyrrolidinopyridine (100 mg) in CH 2

CI

2 and then deprotected in aqueous AcOH (25 mL) to afford 1.30g (81 of2-(trimethylsilyl)ethyl 2-deoxy-3-0- [(R)-3-dodecanoyloxytetradecanoyl]-2-(2,2, 2 -trichloroethoxycarbonylamino)-p-Dglucopyranoside as an amorphous solid: 'H NMR(CDCl 3 5 0.00 9 0.88 8 H), 1.25 28 1.59 4 2.30 2 H, J= 7.5 Hz), 2.52 2 3.42 1 H), 3.55 1 3.66 1 3.83 (dd, 1 H, J= 11.8, 4.6 Hz), 3.94 2 4.57 1 H, J= 8.2 Hz), 4.71 2 5.07 2 5.27 1 H, J= 8.8 Hz).

In the same manner as described in Example the compound prepared in above (1.30 g, 1.51 mmol) was treated with 2,2,2-trichloro-l,1dimethylethyl chloroformate (398 mg, 1.66 mmol) and pyridine (0.15 mL, 1.83 mmol) in CH 2 C1 2 (25 mL) followed by triethylamine (0.42 mL, 3.02 mmol), diphenyl chlorophosphate (0.47 mL, 2.27 mmol) and 4-pyrrolidinopyridine (100 mg) to afford 1.39 g (71 of 2-(trimethylsilyl)ethyl 2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3dodecanoyloxytetradecanoyl]-6--(2,2,2-trichloro-1,1 -dimethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylamino)-p-D-glucopyranoside as an amorphous solid: 'H NMR

(CDC

3 8 0.0 9 0.88 8 1.1 1.7 46 1.77 3 1.85 3 H), 2.23 6 3.34 1 3.59 1 3.80 1 3.96 1 4.32 2 H), 4.63 2 4.83 1 H, J= 11.9 Hz), 5.02 1 H, J= 8.2 Hz), 5.20 1 5.65 2 7.29 10 H).

The compound prepared in above (1.30 g, 1.0 mmol) in CH 2 C1 2 mL) was treated at 0 oC with TFA (5 mL) and then allowed to warm to room temperature for 18 h. The solvent was removed in vacuo and the remaining TFA was removed by azeotroping with toluene. The lactol was treated with the Vilsmeier reagent prepared from DMF (0.39 mL, 5.0 mmol) and oxalyl chloride (0.22 mL, 2.5 mmol) in CH 2

C

2 mL) at 0 oC. The reaction was allowed to warm slowly to room temperature overnight and was partitioned between 50 mL of saturated aqueous NaHCO 3 and ether (50 mL).

WO 98/50399 WO 9850399PCTIUS98/09385 The layers were separated and the organic phase was dried over Na-'SO 4 and concentrated in vacuo. Purification by flash chromatography on silica gel with 10 EtOAc hexanes afforded 1.09 g (90 of 2 -deoxy-4-O-diphenylphosphono3.0-[(R)-3 dodecanoyloxytetradecanoyl]-6-(2,2,2tricloro. 1, 1 -dimethylethoxycarbonyl)2(22,2 trichloroethoxycarbonylamino)-a-D-glucopyranosyl chloride as a white foam: 'H NMR (CDCl 3 8 0.88 6 H, J 6.8 Hz), 1.2 1.70 46 1.78 3 1.88 3 2.18 2 H, J 7.7 Hz), 2.43 (in, 2 4.30 (in, 4 4.72 (in, 3 H),,5.09 (in, I 5.50 (t, 1 H, J= 9.5 Hz), 5.79 1 H, J= 8.0 Hz), 6.27 I H, J= 3.6 Hz), 7.19 (mn, 10 H).

To a solution of compounds prepared in and (540 mg, 0.90 minol, and 1.0 g, 0.82 mmol, respectively) in 1 ,2-dichloroethane (20 mL), powdered 4A molecular sieves (300 ing) were added and the suspension was stirred for 30 min. AgOTf (1.16 g, 4.5 inmol) was added in one portion, after 30 min the slurry was filtered through silica gel and eluted with 30 EtOAc hexanes to afford 1.10 g (75 of dodecanoyloxytetradecanoyl] [2-deoxy-4-0-diphenylphosphono-3 dodecanoyloxytetradecanoyl6o0(222.ticboro- 1, 1 -dimethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylainino)-.Dglucopyranosyl].Lserine benzyl ester: 'H NMR (CDCl 3 5 0.88 12 H, J= 6.5 Hz), 1. 1 1.65 (in, 92 1.77 3 1.85 3 2.1 2.5 (mn, 8 3.67 (in, 2 4.30 (in, 3 4.72 (in, 5 5.18 (mn, 4 5.46 (in, 1 H), 6.07 (in, 1 6.62 I H, J 7.9 Hz), 7.05 7.45 (mn, 15 H).

In the same manner as described in Example the compound prepared in above (1.0 g, 0.56 iniol) was deprotected with zinc (1.83 g, 28 nimol) and acylated with (R)-3-dodecanoyloxytetradecanoic acid (285 mg, 0.67 inmol) in the presence of EEDQ (185 ing, 0.74 inmol) to afford 420 mng (44 of dodecanoyloxytetradecanoyl] -0-[2-deoxy-4-O-diphenylphosphono-2-.[(R)-3dodecanoyloxytetradecanoylamino] -dodecanoyloxytetradecanoyl] -1-Dglucopyranosyl]-L-serine benzyl ester as an amorphous solid.

In the same manner as described in Example the compound prepared in above (420 mg, 0.24 imol) was hydrogenated in the presence of palladium hydroxide on carbon in EtOH (10 mL) and platinum oxide (400 mg) in EtOH AcOH (10: 1) to afford 240 mg (60 of N-[(J?)-3-dodecanoyloxytetradecanoyl]-o-[2.

deoxy-4-0-phosphono-2- 3 -dodecanoyloxytetradecanoylaino] -3 k. WO 98/50399 WO 9850399PCTIUS98/09385 46 dodecanoyloxytetradecanoy]-r3-Dgucopyranosyl-L-seriflc triethylammoniumn salt as a white powder: mp 181-1 82' C; JR (film) 3289, 2956, 2920, 2851, 1731, 1656, 1557, 1467, 1378, 1182, 1108, 1080, 1052, 852, 721 'H NMR (CDC 3

CD

3 OD) 860.88 18 H, J= 6.7 Hz), 1. 1 1. 7 (in, 123 2.2 -2.7 (in, 12 3.06 6 H, J= 7.2 Hz), 3.35 (in, 1 3.70 (in, 6 3.88 (in, 2 4.20 (mn, 1 4.56 I H, J= 8.1 Hz), 4.59 (br s, 1 5.16 4H); 1 3 CNMR (CDC1 3 6 176.9, 173.3,173.2, 172.7, 169.6,169. 1, 101.5, 74.8, 71.2, 70.9,.69.2, 60.5, 53.1, 51.4, 46.1, 41.5, 41.0, 39.2, 34.3, 34.2, 34.0, 32.0, 29.8, 29.7, 29.4, 29.2, 25.6, 25.3, 25.2, 25.1, 22.7, 14.1, 8.7.

Anal. Calcd. for C 93

H

178

N

3 0 19 P C, 66.04; H, 10.73; N, 2.48; P, 1.83.

Found: C, 66.04; H, 10.73; N, 2.48; P, 1.86.

EXAMPLE 14 (B 13) Preparation of N-[(R)-3-Undecanoyloxytetradecanoyl]-O..[2-deoxy-4-O-phosphono-2- 3 -undecanoyloxytetradecanoylanino]3.o[(R)-3undecanoyoxytetadecanoyl]-3Dglucopyranosyl]-L-serine Triethylammonium Salt (Compound R 1

=R

2

=R

3 =n-

CIO)H

2 1CO, X=Y=O, n=m=pq=0, R 4

=R

5

=R

7

=R

9 R6=CO 2 H, R 8

=PO

3 In the same manner as described in Example L-serine benzyl ester (390 mg, 2.0 inmol) was acylated with (R)-3-undecanoyloxytetradecanoic acid (905 mg, 2.2 mmol) in the presence of EDC-MeI (745 mng, 2.5 inmol) in CH 2 C1 2 to afford 1.08 g (92 of N-[(R)-3-undecanoyloxytetradecanoyl]-L-serine benzyl ester: mp 53-54 'C; 'H NMR (CDCL 3 6 0.88 6 H, J= 6.9 Hz), 1. 1 1.7 (in, 44 2.30 2 H, J= 7.7 Hz), 2.49 2 H, J 5.8 Hz), 3.99 (in, 2 4.6 5 (in, 1 5.19 (in, 3 6.5 8 1 H, J= 6.9 Hz), 7.3 5 (hr s, 5 H).

In the same manner as described in Example the compound prepared in Example (1.0 g, 2.02 inmol) was acylated with undecanoyloxytetradecanoic acid (915 mg, 2.22 inmol) in the presence of EDC-MeI (720 mg, 2.4 inmol) and 4-pyrrolidinopyridine (100 mng) in CH 2 C1 2 and then deprotected in aqueous AcOH (25 mL) to afford 1.41 g (82 of 2-(trimethylsilyl)ethyl 2-deoxy-3-O- -undecanoyloxytetradecanoyl]-2-(2 ,2 ,2-trichloroethoxycarbonylamino)- 1-Dglucopyranoside as an amorphous solid: 'H NMR (CDCl 3 6 0.00 9 0.88 (in, 8 H), 1.25 (in, 32 1.60 (in, 4 2.31 2 H, J 7.5 Hz), 2.52 (in, 2 3.42 (in, 1 H), WO 98/50399 WO 9850399PCT/US98/09385 47 5 (in, I 3.66 (in, 1 3.83 (dd, 1 H, J= 11. 8, 4.6 Hz), 3.94 (in, 2 4.5 7 I 8.2 Hz), 4.71 (in, 2 5.07 (in, 2 5.27 1 H, J 8.7 H-z).

In the same manner as described in Example the compound prepared in above (1.30, 1.53 minol) was treated with 2 ,2,2-trichloro-1,1dimethylethyl chioroformate (403 mg, 1.68 mmol) and pyridine 15 mL, 1.85 rnmol) in CHC1 2 (25 mL) followed by triethylamine (0.43 mL, 3.06 inmol), diphenyl chiorophosphate (0.48 mL, 2.30 mmol) and 4 -pyrrolidinopyridine (100 mg) to afford 1.37 g (70 of 2-(triinethylsilyl)ethyl 2-deoxy-4-O-diphenylphosphono3.0[(R)-3 undecanoyloxytetradecanoyl]-6-0(2,2,2-tncfloro 1,1 -diinethylethoxycarbonyl)-2.(22,2 trichloroethoxycarbonylanino)-13-D-glucopyranoside as an amorphous solid: 'H NMR (CDC1 3 6 0.0 9 0.88 (in, 8 1.1 1.7 (in, 44 1.80 3 1.89 3 H), 2.23 (in, 6 3.58 (in, 3 4.32 (in, 1 4.71 (mn, 2 4.83 I H, J= 12.1 Hz), 5.01 I H, J= 8.1 Hz), 5.20 (in, 1 5.62 (in, 2 7.25 (in, 10 H).

In the same manner as described in Example 1 the compound prepared in above (1.28 g, 1.0 rumol) was deprotected with TFA (5 inL) and then treated with the Vilsineier reagent generated from DMF (0.39miL, 5.0 miol) and oxalyl chloride (0.22 mL, 2.5 inmol) to give 1. 12 g (93 of 2 -deoxy-4-O-diphenylphosphono- 3 3 -un d ec an oyIo x yt e trad e c a n oy I-6- 0 t r ich o ro-I ,1I dimethylethoxycarbonyl)-2-(2,2,2.trichoroethoxycarbonylamnino)a-D-gucopyranosy chloride as a white foam: 'H NMR (CDCI 3 6 0.8 8 6 H, J 6.7 Hz), 1. 1 1. 55 (in, 44 1.78 3 1.88 3 2.18 (in, 2 2.43 (mn, 2 4.34 (in, 4 4.72 (in, 3 5.09 (in, 1 5.50 1 H, J 9.6 Hz), 5.80 I H, J =8.0 Hz), 6.26 I H, J= 3.4 Hz), 7.26 (in, 10 H).

In the same manner as described in Example 13 compounds prepared in and above (530 ing, 0.90 inmol, and 1.0 g, 0.83 iniol, respectively) were coupled in the presence of AgOTf (1 .16 g, 4.5 iniol) to afford 1.11 g (76 of undecanoyloxytetradecanoyl]-O-[12-deoxy-4--diphenylphosphono.3 undecanoyloxytetradecanoyl]-6-O-(22,2trichoro.1,1 -dimethylethoxycarbonyl)-2-(2,2,2nrchloroethoxycarbonylamino).3D-glucopyranosyl].L-.serine benzyl ester: 'H NMR (CDCl 3 6 0.88 (in, 12 1.0 1.65 (in, 88 1.77 3 1.85 3 2.1 2.5 (in, I- WO 98/50399 PCTIUS98/09385 48 8 3.37 (in, 1 3.64 (in, 1 3.85 (mn, 1 4.30 (in, 3 4.78 (in, 5 5.18 (mn, 4 5.46 (in, 1 6.07 (in, I 6.62 1 H, J =7.7 Hz), 7.05 7.45 (in, 15 H).

In the same manner as described in Example the compound prepared in above (1.0 g, 0.57 inmol) was deprotected with zinc (2.0 g, 30.5 iniol) and acylated with (R)-3-undecanoyloxytetradecanoic acid (280 mg, 0.68 mmol) in the presence of EEDQ (185 ing, 0.75 mmol) to afford 470 mng (50 of undecanoyloxytetradecanoyl]-0- [2-deoxy-4-0-diphenylphosphono-2-.[(R)-3 undecanoyloxytetradecanoylamino] -undecanoyloxytetradecanoyl] -o-Dglucopyraniosyl]-L-serine benzyl ester as an amorphous solid.

In the same manner as described in Example the compound prepared in above (470 mg, 0.27 inmol) was hydrogenated in the presence of palladium hydroxide on carbon in EtOH (10 mL) and plat *inum oxide (400 ing) in EtOH AcOH (10: 1) to afford 130 mg (30 of N-[(R)-3-undecanoyloxytetradecanoyl]..o-[2deoxy-4-0-phosphono-2-[(R).3 -undecanoyloxytetradecanoylamino]- 3-0- undecanoyloxytetradecanoyll-3-D-glucopyranosyl]..L-serine triethylaminonium. salt as a white powder: mp 181-183 0 C; IR(filmn)3294,2923, 2853, 1734, 1655, 1466, 1377, 1163, 1080, 721 cin-'; 11H NMR (CDC1 3

CD

3 OD) 6 0.88 18 H, J= 6.8 Hz), 1.1 1.7 (in, 117 2.2 -2.7 (in, 12 3.06 6 H, J= 7.1 Hz), 3.4 -3.2 (in, 5 3.6 -3.9 (in, 4 4.20 1 H, 9.8 Hz), 4.54 1 H, J 8.0 Hz), 4.62 (br. s, I 5.17 (in, 4 I 3

C

NMR (CDCl 3 6 173.5, 173.3, 172.8, 172.2, 169.6, 169.1, 101.5, 77.2, 74.8, 70.9, 69.2, 60.5, 58.5, 53.1, 51.5, 46.1, 41.5, 41.1, 39.2, 34.6, 34.4, 34.1, 32.0, 29.8, 29.7, 29.4, 29.2, 25.6, 25.2, 25.1, 22.7, 18.5, 14.2, 8.7.

Anal. Calcd. for C 90

HI

72

N

3 01 9 P: C, 66.26; H, 10.63; N, 2.58; P, 1.90. Found: C, 66.56; H, 10.57; N, 2.47; P, 1.91.

I- WO 98/50399 WO 9850399PCTIUS98/09385 49 EXAMPLE 15 (B 14) Preparation of N- -Decanoyloxytetradecanoyl]-o-[2.

deoxy-4-O-phosphono-2- 3 -decanoyloxytetradecanoylamino]-3 decanoyloxytetradecanoyl]- p-D-glucopyranosyl]-D-serine Triethylanmmonium, Salt (Compound

R,=R

2

=R

3 =n-C 9 Hl 9 C0, X=Y=O, n=m=p=q=0,

R

4

=R

5

=R

7

=R

9

=H,

R

6

=CO

2 H, R 8 =P0 3

H

2 In the same manner as described in Example D-serine benzyl ester (390 mg, 2.0 mmol) was acylated with 3 -decanoyloxytetradecanoic acid (875 mg, 2.2 mmol) in the presence of EDC-MeI (745 mg, 2.5 mmnol) in CH 2 CI, to afford 1.05 g (91 of N-[(R)-3-decanoyloxytetradecanoyl].D..serine benzyl ester: mp 51-52 IC; 1H NMR (CDC1 3 8 0.88 (in, 6 1. 1 1.7 (in, 34 2.30 2 H, J 7.7 Hz), 2.50 (in, 2 H),3.68 1 3.93 2 H, J 3.1 Hz), 4.62 (mn, I 5.22 (in, 3 6.63 I H, J =6.9 Hz), 7.35 (br s, 5 H).

In the same manner as described in Example the compound prepared in Example (1.0 g, 2.02 mmol) was acylated with decanoyloxytetradecanoic acid (884 mg, 2.22 mmol) in the presence of EDC-Mel (720 mg, 2.4 minol) and 4-pyrrolidinopyridine (100 mg) in CHC1 2 and then deprotected in aqueous AcOH (25 mL) to afford 1 .30g (77 of 2-(triinethylsilyl)ethyl 2-deoxy-3-O- 3 decanoyl oxytetradecanoylI] 2 2 -trichl oro ethoxycarbonyamino) P Dglucopyranoside as an amorphous solid: 'H NMR (CDCl 3 8 0.00 9 0.88 (in, 8 1.25 (mn, 30 1.59 (in, 4 2.30 2 H, J= 7.5 Hz), 2.52 (in, 2 3.42 (in, 1 H), 5 (in, 1 3.66 (in, 1 3.8 3 (dd, I H, J 11. 8, 4.6 Hz), 3.94 (in, 2 4.5 7 I H, J 8.2 Hz), 4.71 (in, 2 5.07 (in, 2 5.27 1 H, J 8.8 Hz).

In the same manner as described in Example the compound prepared in above (1.25g, 1.50 mmol) was treated with 2,2,2-trichloro-1,1dimethylethyl chloroforinate (396 mg, 1.65 minol) and pyridine (0.15 mL, 1.81 inmol) in CH 2 C1, (25 mL) followed by triethylamnine (0.42 mL, 3.00 mmol), diphenyl chlorophosphate (0.47 inL, 2.25 minol) and 4-pyrrolidinopyridine (100 mg) to afford 1.31 g (69 of 2-(triinethylsilyl)ethyl 2-deoxy-4-O-diphenylphosphono-3-O-[(R).3 decanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro- 1, 1 -dimethylethoxycarbonyl) trichloroethoxycarbonylamino)-I3-D-glucopyranoside as an amorphous solid: 'H NMR (CDCl 3 850.0 9 0.89 (in, 8 1. 1 1.7 (in, 34 1.82 3 1.90 3 H), WO 98/50399 PCT[US98/09385 2.30 (in, 4 3.40 1 H, J= 9.6 Hz), 3.65 (mn, 1 3.89 (in, 1 4.32 (in, 2 4.63 (m,2 4.82 I H, J =12.1 Hz), 5. 01 1 H, J =8.2 Hz), 5.63 (in, 2 7.29 (in,

H).

In the same manner as described in Example 1 the compound prepared in above (1.27 g, 1.0 inmol) was deprotected with TFA (5 mL) and then treated with the Vilsmeier reagent generated from DMF (0.39 mL, 5.0 mn-ol) and oxalyl chloride (0.22 mL, 2.5 inmol) to give 1.06 g (89 of 2 3 0- -3 -d e can oyIo x yt et r ade ca no yI] 6 2 2 -tr ic hlIoro- 1,1I dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylaiino)-.-D-glucopyranosyI chloride as a white foam: 'H NMR (CDC 3 6 0.88 6 H, J 6.6 Hz), 1. 1 1.55 (in, 34 1.78 3 1.88 3 2.18 2 H, J= 7.7 Hz), 2.43 (in, 2 4.32 (mn, 4 H), 4.71 (in, 3 4.83 (in, 3 5.09 (in, 1 5.50 I H, J =9.5 Hz), 5.77 1 H, J= Hz), 6.26 I H, J 3.4 Hz), 7.20 (in, 10 H).

In the same manner as described in Example 1 compounds prepared in and above above (520 mg, 0.90 minol, and 1.0 g, 0.84 iniol, respectively) were coupled in the presence of AgOTf 16 g, 4.5 inmol) to afford 1. 13 g (78 of decanoyloxytetradecanoyl] [2-deoxy-4-0-diphenylphosphono-3-o.. decanoyloxytetradecanoyl]-6-0-(2,2,2-trichloro- 1,1 -dimethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylanino)-I3-D-glucopyranosyl].D-serine benzyl ester: 'H NMR (CDCl 3 860.88 12 H, J 6.6 Hz), 1. 1 1.65 (in, 68 1.82 3 1.89 3 2.2 2.6 (in, 8 3.40 (in, I 3.64 (in, I 4.01 (in, 2 4.27 (in, 2 4.44 I H, J 7.1 Hz), 4.60 (in, 2 4.77 (in, 2 5.19 (in, 6 6.61 1 H, J= 8.3 Hz), 7.05 7.45 (in, 15 H).

In the same manner as described in Example the compound prepared in above (1.0 g, 0.58 inmol) was deprotected with zinc (1.9 g, 29 mmiol) and acylated with (R)-3-decanoyloxytetradecanoic acid (280 ing, 0.70 inmol) in the presence of EEDQ (190 ing, 0.77 inmol) to afford 420 mg (44 of decanoyloxytetradecanoy]-0-deoxy-4-0-diphenylphosphono-x. decanoy loxytetradecanoyl amino] -3 0 -dec anoy loxytetradecan oy -Dglucopyranosyl]-D-serine benzyl ester as an amorphous solid.

I. WO 98/50399 WO 9850399PCTIUS98/09385 51 In the same manner as described in Example the compound prep~ared in above (420 mg, 0.25 inmol) was hydrogenated in the presence of palladium hydroxide on carbon in EtOH (10 mL) and platinum oxide (400 mg) in EtOH /AcOH (10:1) to afford 118 mng (30 of 3 -decanoyoxytetradecanoyl]yO-2.

deoxy- 4 phosphono-2 decanoyloxytetradecanoyl amino]-3 decanoyloxytetradecanoyl]-3D-glucopyranosy].Dserine triethylaminonium. salt as a white powder: mp 179-181 0 C; JR (film) 3283, 3100, 2921, 2852, 1732, 1660, 165 1, 1564, 1556, 1464, 1417, 1378, 1322, 1181, 1061, 856, 722 ciw'; 11- NMR (CDC1 3

CD

3 OD) 50.88 18 H,J =6.8 Hz), 1. 1- 1.7 I111 2.2 -2.7 12 3.06(mn, 6 3.33 (mn, 5 3.78 (in, 2 3.95 (in, 2 4.22 (in, 1 4.45 1 H, J= Hz), 4.68 (br. s, 1 5.13 (in, 3 5.26 (in, 1 3 CNMR (CDC1 3 668 173.7, 173.5, 173.1, 171.1, 169.9, 100.3, 75.1, 73.9, 71.9, 71.1, 70.9, 70.2, 60.9, 53.9, 52.7, 46.0, 41.3, 40.8, 39.4, 34.6, 34.4, 31.9, 29.8, 29.7, 29.5, 29.4,25.6,25.4, 25.2, 25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for C 87

H,

66

N

3 0, 9 p: C, 65.75; H, 10.53; N, 2.64; P, 1.95. Found: C, 65.32; H, 10.28; N, 2.53; P, 1.89.

EXAMPLE 16 Preparation of of N-[(R)-3-Decanoyloxytetradecanoyly-o..2.deoxy-4-0ophosphn..2- 3 -decanoyloxytetradecanoylainino]-3 3 -decanoyloxytetradecanoyl]

D

glucopyranosyl]-L-serine Triethylammoniuin Salt. (Compound 3 =n-

C

9

H,

9 C0, X=Y=O, n=in=p=q=0, R,=R 5

=R

7

=R

9 R6=COH, R 8 =P0 3

H

2 In the same manner as described in Example L-serine benzyl ester (250 mng, 1.08 inmol) was acylated with (R)-3-decanoyloxytetradecanoic acid (478 mg, 1.2 inmol) in the presence of EDC-MeI (357 mg, 1.2 mmcl) in CH 2

CI

2 to afford 0.52 g (84 of N-[(R)-3-heptanoyloxytetradecanoyl]yLserine benzyl ester: mp 52-53 OC; 'H NMR (CDC1 3 6 0.87 6 H, J= 6.9 Hz), 1. 1 1.7 (mn, 34 2.29 2 H, J 7.5 Hz), 2.49 2 H, J 5.8 Hz), 3.67 1 3.97 (in, 2 4.63 (in, 1 5.19 (in, 3 6.61 1 H, J 7.1 Hz), 7.3 5 (br s, 5 H).

In the same manner as described in Example 1 the compound prepared in above (500 mg, 0.87 inmol), and the compound prepared in Example (1.08 g, 0.90 inmol) were coupled in the presence of AgOTf 16 g, 4.5 inmol) to afford 1.35 g (89 of 3 -decanoyloxytetradecanoyly-O.[2.deoxy-4-0-.

WO 98/50399 WO 9850399PCT[US98/09385 52 diphenylphosphono-3 -decanoyloxytetradecanoy]60(222trichloro- 1,1dintyehxcroy)2(,,-rclrehxcroyann)pDguoyaoy] L-serine benzyl ester: 'H NMR (CDC1 3 8 0.88 12 H, J= 6.6 Hz), 1.0 1.65 (in, 68 H), 1.77 3 1.85 3 2.1 -2.5 (in, 8 3.38 I H, J= 9.1 Hz), 3.65 (in, 1 H), 3.84 (in, I 4.27 (in, 3 4.70 (in, 5 4.84 (in, 4 5.14 (in, 3 5.46 1 H, J 9.7 Hz), 6.07 (in, 1 6.62 1 H, J 8.0 Hz), 7.05 7.45 (mn, 15 H).

In the same manner as described in Example the compound prepared in above (600 mg, 0.34 inmol) was deprotected with zinc (1.13 g, 17.2 mmol) and acylated with 3 -decanoyloxytetradecanoic acid (15 0 mng, 0. 38 mmol) in the presence of EEDQ (124 mg, 0.50 inmol) to afford 362 mng (60 of decanoyloxytetradecanoyl]-O- [2-deoxy-4-O-diphenylphosphono-2-[(R)-3decanoylIoxytetradecanoyl amino] 3 3 -decanoyloxytetradecano glucopyranosyl]-L-serine benzyl ester as an amorphous solid.

In the same manner as described in Example the compound prepared in above (300 mng, 0.17 minol) was hydrogenated in the presence of palladium on carbon (100 mng) and platinum oxide (200 mng) in THF/AcOH (10: 1) to afford 120 mg (44 of N-[(R)-3-decanoyoxytetradecanoyl]o-[2-deoxy.4-0-.

phosphono-2- -decanoyloxytetradecanoylamino]-.3 decanoyloxytetradecanoyl]-3-D-glucopyranosyl].L-.serine triethylainmoniuin salt as a white powder: mp 175-176' C; JR (film) 3304, 2956, 2923, 2853, 1733, 1654, 154 1, 1466, 1377, 1164, 1107, 1080, 845, 721 cm- 1 'HNMR (CDC 3

CD

3 OD) 5 0.88 18 H,J 6.9 Hz), 1. 1 1. 7 (in, 1 11 2.2 -2.75 (in, 12 3.07 6 H, J= 7.2 Hz), 3.3 7 (in, 1 3.5 3.95 (in, 8 4.21 1 H, 11.0 Hz), 4.54 1 H, J= 8.9 Hz), 4.61 (br.

s, I 5.17 (in, 4 7. 10 1 H, J =9.0 Hz), 7.43 1 H, J 7.9 Hz); 1 3 C NMR (CDCl 3 8 176.3, 173.4, 173.2, 172.8, 172.0, 169.6, 169.2, 101.4, 74.7, 70.9, 69.3, 60.4, 53.2, 51.6,46.1,41.4,41.0,39.1,34.5,34.3,34.2,34.1,31.9,29.8,29.7, 29.6,29.4,29.3, 29.2, 25.5, 25.1, 25.0, 22.7, 14.1, 8.6.

Anal. Calcd. for C 8 7

H,

66

N

3 01 9 P -H 2 0: C, 65.01; H, 10.54; N, 2.61; P, 1.93.

Found: C, 64.92; H, 10.38; N, 2.58; P, 2.06.

WO 98/50399 WO 9850399PCT/US98/09385 53 EXAMPLE 17 (B 16) Preparation ofN-[(R)-3-Nonanoyloxytetradecanoyq..o..[2-deoxy-4-0-phosphono-2.[(R..

3 -nonanoyl oxytetradec anoylIamino] -3 -nonanoyloxytetradecanoyl] -f3-Dglucopyranosyl]-L-serine Triethylamrnonium Salt. (Compound R,=R 2

=R

3 =n-

C

8

H,

7 C0, X=Y=O, n=m=p=q=0, R 4

=R,=R

7

=R

9

R

6

=CO

2 H, R 8 =P0 3

H

2 In the same manner as described in Example L-serine benzyl ester (390 mg, 2.0 mmol) was acylated with 3 -nonanoyloxytetradecanoic acid (780 mg, 2.2 mmol) in the presence of EDC-MeI (845 mg, 2.5 mnmol) in CHC1 2 to afford 1.0 g (89 of N-[(R)-3-nonanoyloxytetradecanoyl]-L-serine benzyl ester: mp 52-53 1H NMR (CDC1 3 8 0.88 6 H, J= 6.6 Hz), 1. 1 1.7 32 2.30 2 H, J= 7.7 Hz), 2.51 2 H, J= 5.8 Hz), 2.62 I H, J= 6.0 Hz), 3.98 (in, 2 4.65 (mn, 1 5.19 (in, 3 6.5 8 1 H, J 6.8 Hz), 7.3 5 (br s, 5 H).

In the same manner as described in Example the compound prepared in Example (1.0 g, 2.02 mmol) was acylated with nonanoyloxytetradecanoic acid (852 mg, 2.22 inmol) in the presence of EDC-MeI (720 mg, 2.4 mmol) and 4-pyrrolidinopyridine (100 ing) in CH 2 and then deprotected in aqueous AcOH (25 mL) to afford 1.31 g (79 of 2-(trimethylsilyl)ethyl 2-deoxy-3-0- 3 -nonanoy Ioxytetradecanoyl 2.-(2,2,2..trichl oro etho xycarbony aino) glucopyranoside as an amorphous solid: 'H NMR (CDCl 3 5 0.00 9 0.88 (in, 8 H), 1.25 (in, 28 1.59 (in, 4 2.30 2 H, J 7.5 Hz), 2.52 (in, 2 3.42 (in, 1 H), 5 (in, 1 3.66 (in, 1 3.8 3 (dd, 1 H, J 11. 8, 4.6 Hz), 3.94 (in, 2 4.5 7 1 H, J 8.2 Hz), 4.71 (in, 2 5.07 (in, 2 5.27 1 H, J 8.8 Hz).

In the same manner as described in Example the compound prepared in above (1.25 g, 1.52 inmol) was treated with 2,2,2-trichioro- 1,1dimethylethyl chloroformate (400 ing, 1.67 mmol) and pyridine 15 inL, 1.84 mmol) in CH 2

CI

2 (25 inL) followed by triethylamnine (0.42 -mL, 3.04 inmol), diphenyl chiorophosphate (0.47 mL, 2.28 inmol) and 4-pyrrolidinopyridine (100 mg) to afford 1.30 g (67 of 2-(triinethylsilyl)ethyl 2-deoxy-4-0-diphenylphosphono-3-0-[(R)-3nonanoyloxytetradecanoyl]-6-0-(2,2,2-trichoro- 1,1 -diinethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylanino)-3-D-glucopyranoside as an amorphous solid: 'H NMR (CDC1 3 8 0.0 9 0.88 (in, 8 1.1 1.7 (in, 32 1.82 3 1.89 3 H), 2.22 (in, 6 3.33 (in, 1 3.53 (in, 1 3.80 (in, 1 3.96 (in, I 4.31 (in, 2 H), WO 98/50399 WO 9850399PCTIUS98/09385 54 4.55 (in,2 4.83 1 H, J= 12.0 Hz), 5.01 1 H, J=7.9 Hz), 5.62(in, 1 7.28 (mn, 10 H).

In the same manner as described in Example 1 the compound prepared in above (1.26 g, 1.0 minol) was deprotected with TFA (5 mL) and then treated with the Vilsmeier reagent generated from DMF (0.39 mL, 5.0 mmol) and oxalyl chloride (0.22 mL, 2.5 mmol) to give 1.07 g (91 of 2 3 0- 3 -n on a n oyIo x yt et ra de c ano yI] 6 2 2, 2 -t r ichlIor o 1,1I dimethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylamino)a-DglucopyranosyI chloride as a white foam: 'H NMR (CDCl 3 6 0.88 6 H, J 6.9 Hz), 1.25 1.5 5 (in, 32 1.78 3 1.88 3 2.18 2 H, J= 7.7 Hz), 2.43 (in, 2 4.34 (in, 4 H), 4.70 (in, 3 4.83 (mn, 3 5.09 (in, 1 5.51 1 H, J= 10.2 Hz), 5.78 1 H, J= 8. 0 Hz), 6.2 5 I H, J= 3.6 Hz), 7.19 (in, 10 H).

In the same manmer as described in Example 1 compounds prepared in and above (505 mg, 0.90 mmol, and 1.0 g, 0.85 minol, respectively) were coupled in the presence of AgOTf(L .16 g, 4.5 inmol) to afford 1.03 g (71 of nonanoyloxytetradecanoyl -deoxy..4..0.diphenylphosphono-3 0- nonanoyloxytetradecanoyl]-6-0-(2,2,2-trichloro. 1, 1 -dimethylethoxycarbonyl)-2-(2,2,2.

trichloroethoxycarbonylamino)-pDgucopyranosy].L-serine benzyl ester: 'H NMR (CDCl 3 6 0.88 12 H, J= 6.9 Hz), 1.0 1.65 (in, 64 1.78 3 1.82 3 2.1 2.5 (in, 8 3.38 (in, 1 3.64 (in, 1 3.83 (mn, 1 4.25 (in, 3 4.73 (mn, 5 H), 5.18 (in, 5 6.07 (mn, 1 6.60 1 H, J 7.8 Hz), 7.05 7.45 (in, 15 H).

In the same manner as described in Example the compound prepared in above (1.0 g, 0.59 minol) was deprotected with zinc (1.93 g, 29.5 inmol) and acylated with (R)-3-nonanoyloxytetradecanoic acid (273 ing, 0.71 niiol) in the presence of EEDQ (195 mg, 0.78 iniol) to afford 405 mng (42 of nonanoyloxytetradecanoyl] -0-[deoxy-4-0-diphenylphosphono.2-.[(R)-3 nonanoyloxytetradecanoylainino] -nonanoyloxytetradecanoyl]-3-Dglucopyranosyl]-L-serine benzyl ester as an amorphous solid.

In the same manner as described in Example the compound prepared in above (405 ing, 0.25 inrol) was hydrogenated in the presence of palladium hydroxide on carbon in EtOH (10 mL) and platinum oxide (400 mng) in EtOH I- WO 98/50399 PCTIUS98/09385 /AcOH (10: 1) to afford 185 mg (48 of 3 -nonanoyloxytetradecanoyl]yO[2.

deaxy-4-O-phosphono-2- -nonano ylIoxytetrade canoy Iamino] nonanoyloxytetradecanoy]-f3-D-glucopyranosyl].L-.serine triethylammoniumn salt as a white powder: mp 177-179' C; IR (film) 3306, 2955, 2923, 2853, 1732, 1660, 1538, 1467, 1378, 1252, 1165, 1106, 1080, 960, 844, 722 1 1- NMR (CDCl 3

CD

3 OD) 8 0.88 18 H, J= 6.8 Hz), 1.1 1.7 (in, 105 2.2 2.75 (in, 12 3.07 6 H, J= 7.1 Hz), 3.2 3.5 (in, 5 3.85 (mn, 4 4.23 I H, 10.2 Hz), 4.51 1 H, J 8.0 Hz), 4.64 (br. s, 1 5.18 (in, 4 I IC NMR (CDC 3 6 173.3, 172.8, 172.2, 169.6, 169. 1, 101.5, 74.8, 70.9, 70.8, 69.3, 60.5, 53.2, 51.5, 46.1, 41.5, 41.0, 39.2, 34.5, 34.3, 34.1, 32.0, 31.9, 29.8, 29.6, 29.4, 29.3, 25.6, 25.2, 25.1, 22.7, 14.1, 8.7.

Anal. Calcd. for C 84

H

16

,N

3 0 19 P: C, 65.21; H, 10.42; N, 2.72; P, 2.00. Found: C, 65.48; H, 10.32; N, 2.62; P, 2.12.

EXAMPLE 18 (B317) Preparation ofN-[(R)-3-Octanoyoxytetradecanoyl-o-[2..deoxy-4..phosphono2[(R).

3 -octanoyloxytetradecanoylamino]-3 -octanoyloxytetradecanoyl] -13-Dglucopyranosyl]-L-serine Triethylainmoniumn Salt (Compound R, =R 2

=R

3 =n-

C

7 HJ1 5 CO, X=Y=O, n=m=p=q=0, R 4

=R

5

=R

7

=R

9 R6=COH, R 8 =p0 3

H

2 In the same manner as described in Example L-serine benzyl ester (390 mg, 2.0 inmol) was acylated with (R)-3-octanoyloxytetradecanoic acid (815 mg, 2.2 inmol) in the presence of EDC-MeI (745 mg, 2.5 nimol) in CH 2 C1 2 to afford 1.02 g (93 ofN-[(R)-3-octanoyloxytetradecanoyl-L.serine benzyl ester: mp, 50-51 'HNMR (CDC1 3 8 0.88 6 H, J= 6.8 Hz), 1. 1 1.7 (in, 30 2.30 2 H, J= 7.7 Hz), 2.51 (d, 2 H, J 5.8 Hz), 2.60 1 H, J 6.0 Hz), 3.97 (in, 2 4.65 (mn, I 5.22 (in, 3 H), 6.61 1 H, J 6.9 Hz), 7.3 5 (br s, 5 H).

In the same manner as described in Example the compound prepared in Example (1.0 g, 2.02 mniol) was acylated with octanoyloxytetradecanoic acid (821 mng, 2.22 inmol) in the presence of EDC-MeI (720 ing, 2.4 inmol) and 4-pyrrolidinopyridine (100 mg) in CH 2

C

2 and then deprotected in 90 aqueous AcOH (25 inL) to afford 1 3 5 g (83 of 2-(trimethylsilyl)ethyl 2-deoxy- 3 0[R--caolxttadcnyl2(,,-rclrotoyabnlmn)pD glucopyranoside as an amorphous solid: 'H NMR (Ct)C1 3 6 0.00 9 0.88 (mn, 8 I- WO 98/50399 WO 9850399PCTIUS98/09385 56 1.25 (in, 26 1.60 (in, 4 2.30 2 H, J= 7.5 Hz), 2.53 (mn, 2 3.42 (in, 1 H), 3.53 (in, I 3.66 (in, 1 3.83 (dd, I H, J =11 4.4 Hz), 3.94 (mn, 2 4.56 I H, J= 8.3 Hz), 4.64 I H, J= 11.8 Hz), 4.77 1 H, J= 11.8 Hz), 5.08 (mn, 2 5.30 (br. s, 1 H).

In the same manner as described in Example the compound prepared in above (1.30 g, 1.61 inmol) was treated with 2,2,2-trichloro-1,1..

dimethylethyl chioroformate (425 mg, 1.77 inmol) and pyridine 16 mL, 1.95 minol) in CH 2 Cl 2 (25 inL) followed by triethylainine (0.45 mL, 3.22 mmol), diphenyl chiorophosphate (0.50 mL, 2.42 mmol) and 4-pyrrolidinopyridine (100 mg) to afford 1.42 g (71 of 2-(trimethylsilyl)ethyl 2-deoxy-4-O-diphenylphosphono3.0-[(R)-3 octanoyloxytetradecanoyl]-6-O-(2,2,2-trichloro-.1,1I -dimethylethoxycarbony1)-2-(2,2,2.

trichloroethoxycarbonylainino)-p-D-glucopyranoside as an amorphous solid: 'H NMR (CDC1 3 6 0.0 9 0.88 (mn, 8 1.1 1.7 30 1.82 3 1.89 3 H), 2.23 (in, 6 3.37 (in, 1 3.65 (in, 1 3.83 (in, I 3.96 (in, 1 4.55 (in, 2 H), 4.83 1 H, J= 11. 8 Hz), 5.01 I H, J= 8.2 Hz), 5.20 (in, 1 7.29 (in, 10 H).

In the same manner as described in Example 1 the compound prepared in above (1.24 g, 1.0 inmol) was deprotected. with TFA (5 mL) and then treated with the Vilsineier reagent generated from DMF (0.39 inL, 5.0 mmol) and oxalyl chloride (0.22 mL, 2.5 inmol) to give 1.0 g (87 of 2 -deoxy-4-O-diphenylphosphono- 3--()3otnyoxttaeaol 10(,,-rclr-,1 -diinethylethoxycarbonyl)- 2 2 2 2 -trichloroethoxycarbonylamino)-a-D-glucopyranosyI chloride as a white foam: NMR (CDCl 3 6 0.88 6 H, J= 6.7 Hz), 1.25 1.55 (mn, 30 1.78 3 1.88 3 2.18 2 H, J= 7.7 Hz), 2.43 (in, 2 4.29 (in, 4 4.72 (in, 3 5.09 (in, I 5.51 1 H, J =9.9 Hz), 5.79 1 H, J =7.9 Hz), 6.25 I H,J =3.5 Hz), 7.29 In the same manner as described in Example 1 compounds prepared in and above (490 mg, 0.90 imol, and 1.0 g, 0.86 iniol, respectively) were coupled in the presence of AgOTf .16 g, 4.5 inmol) to afford 0.99 g (69 of octanoyloxytetradecanoyl]. 0- [2-deoxy-4-O-diphenylphosphono.3 octanoyloxytetradecanoyly6.0-(2,2,2.trichloro- 1, 1 -dimnethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylamino).p3D-glucopyranosyl].L-serine benzyl ester: 'H NMR WO 98/50399 WO 9850399PCTIUS98/09385 57 (CDCl 3 6 0.88 12 H,J= 6.9 Hz), 1.0 1.65 (in, 60 1.77 3 1.85 3 2.1 2.Z5 (in, 8 3.37 (in, 1 3.65 (in, 1 3.83 (mn, 1 4.27 (in, 3 4.72 (in, 5 H), 5.18 (mn, 4 5.46 1 H, J 9.8 Hz), 6.06 (in, I 6.60 1 H, J 8.0 Hz), 7.05 7.45 (in, 15 H).

In the same manner as described in Example the compound prepared in above (0.95 g, 0.57 mmol) was deprotected with zinc (1.86 g, 28.5 inmol) and acylated with (R)-3-octanoyloxytetradecanoic acid (252 mg, 0.68 inmol) in the presence of EEDQ (185 mg, 0.75 inmol) to afford 433 mg (47 of octanoyloxytetradecanoyl] -0-[2-deoxy-4-0-diphenylphosphono.2.[(R)-3 octanoyloxytetradecanoyIamino] 3- 0- 3 octanoylIoxytetrad ecanoyl~~.

glucopyranosyl]-L-serine benzyl ester as an amorphous solid.

In the same manner as described in Example the compound prepared in above (433 mng, 0.27 inmol) was hydrogenated in the presence of palladium hydroxide on carbon (250 ing) in EtOH (10 m1L) and platinum oxide (400 mg) in EtOH AcOH (10: 1) to afford 196 mng (48 of 3 -octanoyloxytetradecanoyl].

0- [2-deoxy-4-0-phosphono-2- -octanoyloxytetradecanoylainino-3 octanoyloxytetradecanoy]-3-D-glucopyranosyl]-L-.serine triethylammoniuin salt as a white powder: mp, 177-1781 C; IR (film) 3296, 2956, 2923, 2853, 1732, 1645, 1546, 1466, 1378, 1315, 1170, 1082, 1056, 961, 846,722 1 H NMR (CDC1 3

CD

3 OD) 6 0.88 18 H, J= 6.6 Hz), 1.1 1.7 (in, 99 2.2 2.75 (in, 12 3.08 6 H, J= 7.1 Hz), 3.3 9 I H, J 8.8 Hz), 3.6 4. 0 (in, 8 4.22 I H, 10. 3 Hz), 4.5 3 1 H, J Hz), 4.63 (in, 1 5.18 (mn,4H), 7.04 1 H, J =8.8 Hz), 7.42 1 H, J Hz); 1 3 C NMR (CDCI 3 6 176.8, 173.3, 173.2, 172.7, 172.2, 169.6, 169.1, 101.5, 74.8, 70.9, 70.8, 69.3, 60.5, 53.2, 51.5, 46.2, 41.5, 41.1, 39.2, 34.5, 34.3, 34.1, 34.0, 32.0, 31.8, 29.8, 29.6, 29.4, 29.3, 29.2, 29.1, 25.6, 25.3, 25.2, 25.0, 22.7, 14.1, 8.7.

Anal. Calcd. for C 8 1 1 54

N

3 01 9 P -H 2 0: C, 63.87; H, 10.32; N, 2.76; P, 2.03.

Found: C, 63.96; H, 10.29; N, 2.69; P, 1.67.

WO 98/50399 PCTIUS98/09385 58 EXAMPLE 19 (B 18) Preparation ofN-[(R)-3-Heptanoyloxytetradecanoyl].O[2'.deoxy-4-0O.phosphono.2-(R)- 3 -heptanoyloxytetradecanoylamino]-3 -heptanoyloxytetradecanoyl].j3Dglucopyranosyl]-L-serine Triethylammonium. Salt (Compound R,=R 2

=R

3 =n-

C

6 H,1 3 C0, X=Y=O, n=m=p=q=0, R,=R 5

=R,=R

9

R

6

=CO

2 H, R 8 =P0 3

H

2 In the same manner as described in Example L-serine benzyl ester (390 mg, 2.0 mnmol) was acylated with (R)-3-heptanoyloxytetradecanoic acid (780 mg, 2.2 mmol) in the presence of EDC-MeI (745 mg, 2.5 mmol) in CH 2 Cl, to afford 0.97 g (91 of N-[(R)-3-heptanoyloxytetradecanoyl)-L-serine benzyl ester: mp 46-48 IC; 'H NMR (CDCl 3 5 0.88 6 H, J= 6.9 Hz), 1.1 1.7 (in, 28 2.30 2 H, J= 7.7 Hz), 2.50 2 H, J= 5.8 Hz), 2.62 1 H, J= 6.0 Hz), 3.97 (in, 2 4.65 (in, 1 5.19 (in, 3 6.61 1 H, J =6.9 Hz), 7.3 5 (br s, 5 H).

In the same manner as described in Example the compound prepared in Example (1.0 g, 2.02 nol) was acylated with heptanoyloxytetradecanoic acid (790 mg, 2.22 minol) in the presence of EDC-MeI (720 mg, 2.4 mmol) and 4-pyrrolidinopyridine (100 mg) in CH,C1 2 and then deprotected in aqueous AcOH (25 mL) to afford 1.30g (81 of 2-(triinethylsilyl)ethyl 2-deoxy- -heptanoyloxytetradecanoyl] 2 2 2 2 -trichloroethoxycarbonylanino)-.WDglucopyranoside as an amorphous solid: 'H NMR (CDCl 3 )850.00 9 0.88 (in, 8 1.25 (in, 24 1.59 (in, 4 2.30 2 H, J= 7.5 Hz), 2.52 (in, 2 3.42 (in, 1 H), 5 (in, I 3.66 (in, I 3.83 (dd, I H, J =11. 5, 4.2 Hz), 3.94 (in, 2 4.5 7 1 H,J =8.3 Hz), 4.64 1 H, J= 12.1 Hz), 4.76 1 H, J 11.9 Hz), 5.09 2H), 5.31 1 H, J= 8.7 Hz).

In the same manner as described in Example the compound prepared in above (1.25g, 1.58 inmol) was treated with 2,2,2-trichloro-1,1dimethylethyl chioroformate (417 mg, 1.74 inmol) and pyridine 15 m.L, 1.91 inmol) in CH 2

CI

2 (25 mL) followed by triethylamine (0.44 niL, 3.16 inmol), diphenyl chiorophosphate (0.49 mL, 2.37 inmol) and 4-pyrrolidinopyridine (100 mg) to afford 1.34 g (69 of 2-(trimethylsilyl)ethyl 2-deoxy-4-O-diphenylphosphono-3-O-[(R)-3heptanoyloxytetradecanoyl]-6-O-(2,2,2.trichlro. 1, 1 -dimethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylamino)-3-D-glucopyranoside as an amorphous solid: 'H NMR (CDCl 3 6 0.0 9 0.88 (in, 8 1.1 1.7 (in, 28 1.82 3 1.89 3 H), WO 98/50399 PCTIUS98/09385 59 2.35 (in, 4 3.37 (in, I 3.61 (mn, I 3.80 (in, 1 4.32 (in, 2 4.63 (in, 2 H), 4.83 1 H, J 12.0 Hz), 5.01 I H, 8.2 Hz), 5.62 (in, 2 7.29 (in, 10 H).

In the same manner as described in Example the compound prepared in above (1.23 g, 1.0 innol) was deprotected with TFA (5 mL) and then treated with the Vilsmeier reagent generated from DMF (0.39 mL, 5.0 minol) and oxalyl chloride (0.22 inL, 2.5 minol) to give 1.0 g (87 of 2 -deoxy-4-O-diphenylphosphono.

3 3 -h ept an oyIo x y tet radcanoylI 6 2 2 ,2 -tri chIo ro 1,1I dimethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylamino)-a-DglucopyranoyI chloride as a white foam: 'H NMR (CDCl 3 8 0.88 6 H, J 6.9 Hz), 1.25 1.55 (mn, 28 1.78 3 1.88 3 2.18 2 H, J= 7.6 Hz), 2.43 2 4.26 (mn,4 H), 4.73 (in, 3 5.09 (in, I 5.51 1 H, J =10.2 Hz), 5.77 I H, J 8.0 Hz), 6.25 1 H, J =3.3 Hz), 7.19 (mn, In the same manner as described in Example 1 compounds prepared in and above (480 ing, 0.90 inmol, and 0.98g, 0.86 minol, respectively) were coupled in the presence of AgOTf( 16 g, 4.5 inmol) to afford 1.06 g (75 of heptanoyloxytetradecanoyl]-O-[2-deoxy-4..diphenyl phosphono-3 heptanoyloxytetradecanoyl]-6-0.(2,2,2trichloro.1,1 -diinethylethoxycarbonyl)-2-(2,2,2.

trichloroethoxycarbonylainino)-f.D-glucopyranosyl]Lserine benzyl ester: 'H NMR (CDCl 3 )8 0.88 (in, 12 1.0 1.65 (in, 56 1.77 3 1.85 3 2.1 2.5 (in, 8 3.38 (in, 1 3.64 (in, I 3.83 (in, I 4.25 (in, 3 4.78 (mn, 5 5.16 (in, 4 5.46 1 H, J 9.9 Hz), 6.06 (in, 1 6.60 I H, J 7.7 Hz), 7.05 7.45 (in,

H).

In the same manner as described in Example the compound prepared in above (1.0 g, 0.61 mniol) was deprotected with zinc (2.0 g, 30.5 inmol) and acylated with (R)-3-heptanoyloxytetradecanoic acid (260 ing, 0.73 iniol) in the presence of EEDQ (200 nmg, 0.80 iniol) to afford 440 mng (45 of heptanoyloxytetradecanoyl]-O-[2deoxy40diphenylphosphono2[(R)- 3 heptanoyloxytetradecanoylamino]-3-o -heptanoyloxytetradecanoyl]-3-Dglucopyranosyl]-L-serine benzyl ester as an amorphous solid.

In the same manner as described in Example the compound prepared in above (440 ing, 0.28 mmol) was hydrogenated in the presence of WO 98/50399 PCT/US98/09385 palladium hydroxide on carbon (250 mg) in EtOH (10 mL) and platinum oxide (400 mg) in- EtOH AcOH (10:1) to afford 208 mg (51 of heptanoyloxytetradecanoyl]-O-[2-deoxy-4-O-phosphono-2-[(R)-3heptanoyloxytetradecanoylamino]-3-O-[(R)-3-heptanoyloxytetradecanoyl]-P-Dglucopyranosyl]-L-serine triethylammonium salt as a white powder: mp 176-177° C; IR (film) 3307, 2956, 2924, 2854, 1732, 1650, 1545, 1466, 1378, 1316, 1170, 1080, 956, 841, 722 'HNMR (CDCI 3 CD30D) 5 0.88 18 1.1 1.7 93 2.2 2.75 12 3.08 6 H, J= 7.2 Hz), 3.40 1 H, J= 10.2 Hz), 3.6 4.0 7 H), 4.24 2 4.52 1 H, J= 8.0 Hz), 4.63 1 5.19 4 7.04 1 H, J= 8.6 Hz), 7.40 1 H, J= 8.4 Hz); 3 C NMR (CDC1 3 177.1,173.2,173.1, 172.7, 172.3, 169.5, 168.9, 101.5, 75.0 74.8, 71.2, 70.9, 69.1, 60.5, 53.1, 51.4, 46.1,41.5, 41.0, 39.2, 34.5, 34.3, 34.1,34.0, 31.9, 31.6, 31.5, 29.8, 29.6, 29.4, 29.0,28.9, 28.8, 25.6, 25.3, 25.1, 25.0, 22.7, 22.6, 14.1, 8.7.

Anal. Calcd. for C 78

H,

48

N

3 0 9 C, 64.04; H, 10.20; N, 2.87; P, 2.12. Found: C, 63.77; H, 10.11; N, 2.85; P, 2.02.

EXAMPLE 20 (B19) Preparation of 2-[(R)-3-Tetradecanoyloxytetradecanoylamino]ethyl 2-Deoxy-4-0phosphono-3-O-[(R)-3-tetradecanoyoxytetradecanoyl]-2-[(R)-3tetradecanoyoxytetradecanoylamino]-P-D-glucopyranoside Triethylammonium Salt (Compound R,=R 2

=R

3 =n-C, 3

H

27 CO, X=Y=O, n=m=p=q=0, R 4

=R

5

=R,=R

7

=R

9

=H,

R

8

=PO

3

H

2 2-Amino-1 -(t-butyldiphenylsilyloxy)ethane (330 mg, 1.1 mmol) and 3-tetradecanoyloxytetradecanoic acid (500 mg, 1.1 mmol) were dissolved in CH 2 Cl 2 mL) and treated with powdered 4 A molecular sieves (500 mg). After 1 h EEDQ (297 mg, 1.2 mmol) was added and the reaction was stirred for 18 h, filtered through Celite® and concentrated in vacuo. The residue was chromatographed over silica gel using EtOAc hexanes to give 675 mg (92 of a colorless solid. A portion of this material (500 mg, 0.68 mmol) was deprotected with TBAF (1 M in THF, 1 mL, 1 mmol) in THF (5 mL) by stirring at room temperature for 2 h. The reaction mixture was diluted with (50 mL) and washed with brine (2 x 50 mL). The brine was back extracted with Et 2 O (2 x 50 mL) and the combined organic extracts were dried over NaSO 4 and WO 98/50399 WO 9850399PCTIUS98/09385 61 concentrated in vacuo to afford 338 mg (62 of tetroadecanoyloxytetradecanoylamino] ethanol as an off-white solid.

In the same manner as described in Example the compound prepared in above (338 mg, 0.68 mmol) and the compound prepared in Example 2-(4) (786 mg, 0.61 mmol) were coupled in the presence of mercury cyanide (770 mg, 3.05 mmol) to give 245 mg of 2 3 -tetradecanoyloxytetradecanoylamino] ethyl 2deoxy-4-O-diphenylphosphono-3-o-[(R).3 -tetradecanoyoxytetradecanoylp6.0-(2,2,2trichioro- 1, 1 -dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-.-Dglucopyranoside as an amorphous solid: 'H NMR (CDC1 3 8 0.88 12 H, J= 6.9 Hz), 1. 1 1.8 (in, 84 1.81 3 1.89 3 2.15 -2.55 (mn, 8 3.25 (in, 1 3.47 (in, 2 3.67 (mn, 1 3.83 (in, 2 4.28 (dd, 1 H, J =12.2, 4.9 Hz), 4.36 1 H, J 11.0OHz), 4.68 (mn,2 4.78 I H, J= 11.6 Hz), 4.94 I H,J= 11.6 Hz), 5.16 (m, 2 5.53 I H, J= 10.0 Hz), 6.06 I H, J= 4.9 Hz), 6.19 (mn, 1 7.25 (in, 10 H).

In the same manner as described in Example the compound prepared in above (500 mg, 0.29 mmol) was deprotected with zinc (980 mg, iniol) and then acylated with (R)-3-tetradecanoyloxytetradecanoic acid (155 ing, 0.34 inmol) in the presence of EEDQ (1 10 mng, 0.44 inmol) to give 315 mg of tetradecanoyloxytetradecanoylamino]ethyl 2-deoxy-4-O-diphenylphosphono-3-o tetradecanoyoxytetradecanoyl] -tetradecanoyoxytetradecanoylainino]

-PD

glucopyranoside as an amorphous solid.

In the same manner as described in Example the compound prepared in above (200 ing, 0. 113 inmol) was hydrogenated in the presence of platinum oxide (100 mg) to give 142 mng (76 of tetradecanoyloxytetradecanoylIamnino] ethylI 2 -deoxy-4-O-phosphono-3 tetradecanoyoxytetradecanoyl] -tetradecanoyoxytetradecanoylamino] -f-Dglucopyranoside triethylaminoniuinsalt as a white solid: mnp 175-176* C; IR (film) 3285, 3098, 2955, 2919, 2851, 1731, 1659, 1642, 1556, 1468, 1379,1250, 1228, 1174, 1110, 1083, 1046, 962, 857 'H NMR (CDC1 3

CD

3 OD) 8 0.88 18 H, J= 6.0 Hz), 1 .1 1. 7 (in, 13 5 2.2 -2.7 (in, 15 3.06 6 H, J= 7.1 Hz), 3.2 -4.1 (in, 8 4.21 (q, 1 H, J= 9.9 Hz), 4.51 1 H, J= 8.2 Hz), 5.05 -5.25 (in, 4 7.33 1 H, 8.5 Hz), 7.50 (brt, 1 H,J= 4.8 Hz); 3

CNMR(CDC

3 )8 173.7,173.3,170.6,170.3,169.9,100.9, WO 98/50399 PCTIUS98/09385 62 75.8, 73.0, 71.3, 71.1, 70.9, 70.6, 68.3, 60.6,55.1, 45.7, 41.6, 41.2, 39.5, 34.6, 34.5, 34.4, 32--0, 29.8, 29.4, 29.3, 25.4, 25.1, 22.7, 14.2, 8.6.

Anal. Calcd. for C 98 H,1 90

N

3 01 7 P -2 H 2 0: C, 67.28; H, 11.18; N, 2.40; P, 1.77.

Found: C, 67.0 1; H, 11. 18; N, 2.15; P, 2.01.

EXAMPLE 21 (B320) Preparation of 2 -[(R)-3-Decanoyloxytetradecanoylamino]ethyl 2 -Deoxy-4-O-phosphono-.

3 -O-[(R)-3-decanoyoxytetradecanoy]-2-[(R).3 -decanoyloxytetradecanoylamino] -p-Dglucopyranoside Triethylammonium Salt (Compound

RI=R,=R

3 ==n-C 9

HJ

9

CO,

X=Y=O, n=m=p=q=0, R 4

=R,=R

6

=R

7

R

8

=PO

3 In the same manner as described in Example 2-amino-1-QIbutyldiphenylsilyloxy)ethane (450 mg, 1.5 mmol) was acylated with decanoyloxytetradecanoic acid (600 mg, 1.5 mnmol) in the presence of EDC-MeI (594 mg, 2.0 mmol) and then deprotected with TBAF (1.0 M in THF, 2.5 mL, 2.5 mmol) in THF (10 mL) to afford 488 mg (81 of decanoyloxytetradecanoylamino]ethanoI as an off-white solid.

In the same manner as described in Example 1 the compound prepared in above (385 g, 0.87 mmol) and the compound prepared in Example 1 5-(4) (1.05 g, 0.87 mmol) were coupled in the presence of AgOTf (560 mg, 2.2 mmol) to give 1.04 g (74 of 2 -[(R)-3-decanoyloxytetradecanoylamino]ethyl 2-deoxy-4-Odiphenylphosphono-3 -decanoyoxytetradecanoyl] ,2,2-trichloro- 1,1Idimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino).i3Dglucopyranoside as an amorphous solid: 'H NMR (CDC 3 50.88 12 H, J=6.9 Hz), 1.1 -1.6(in, 68 1.78 3 1.88 3 2.18 2 H, J 7.7 Hz), 2.44 (in, 2 4.34 (in, 5 H), 4.72 (in, 2 4.83 1 H, J 9.3 Hz), 5.09 (in, 1 5.51 1 H, J =10.2 Hz), 5.79 1 H, J 8.0 Hz), 6.26 1 H, J 3.4 Hz), 7.31 (mn, 10 H).

In the same manner as described in Example the compound prepared in above (700 mg, 0.44 mmol) was deprotected with zinc (1.42 g, 21.7 minol) and then acylated with (R)-3-decanoyloxytetradecanoic acid (190 mg, 0.48 mrnol) in the presence of EEDQ (148 mg, 0.6 mnol) to give 432 mg (62 of decanoyloxytetradecanoylamnino]ethyI 2-deoxy-4-O-diphenylphosphono..3.0-[(R).3 WO 98/50399 WO 9850399PCT/US98/09385 63 decanoyoxytetradecanoyl]-2- -decanoy lox ytetradecanoylai no] -P-D.

glucopyranoside as an amorphous solid.

In the same manner as described in Example the compound prepared in above (400 mg, 0.25 mmol) was hydrogenated in the presence of platinum oxide (200 mg) to give 200 mg (52 of 2 3 -decanoylIoxytetradecaiioylamino] ethyl 2 -deoxy -4 -O-pho sphono,- 3 0- -3 decano yoxytetrad ecan oylIy.-2- -3 decanoyloxytetradecanoylamino]-J3-D-glucopyranoside triethylammonium salt as a white solid: mp 165-166O C; IR (film) 3289, 3094, 2956, 2922, 2853, 1732,1658,1644,1556, 1467, 1379, 1247, 1164, 1107, 1081, 1048 cm'1; 'H NMR (CDCl 3

CD

3 OD) 8 0.88 (t, 18 H, J=6.9 Hz), 1.1 1.7 I111 2.2 -2.7 15 3.05 6H, J=7.1 Hz), 3.2 3.85 (in, 9 4.52 I H, J 8.2 Hz), 5.05 5.25 (in, 4 7.21 1 H, J 8.5 Hz), 7.42 (br t, I 1 3 CNMR(CDC1 3 )8 173.8, 173.3, 170.7, 170.3, 170.0, 100.9, 75.6, 73.0, 71.3,70.9, 70.6,68.3,60.7,55.0,45.8,41.6,41.2,39.5,34.5,34.4,34.1,31.9, 29.8,29.6, 29.5, 29.4, 25.4, 25.1, 22.7, 14.2, 8.6.

Anal. Calcd. for C 86

H,

66

N

3 0, 7 P H 2 0: C, 66.08; H, 10.83; N, 2.69; P, 1.98.

Found: C, 65.80; H, 10.63; N, 2.63; P, 2.04.

EXAMPLE 22 (B21) Preparation of 3- -Tetradecanoyloxytetradecanoylamino]propyl 2-Deoxy-4-0phosp h on o 0 -t e trad e canoy ox yt etrad e can oy I] -2 -3 tetradecanoyloxytetradecanoylamino])-I3-D-glucopyranoside Triethylammonium Salt (Compound R,=R 2

=R

3 =n-C1 3 H1 27 C0, X=Y=O, n= 1, m=p=q=0, R 4

=R

5

=R

6

=R

7

=R

9

=H,

R

8 =P0 3

H

2 In the same manner as described in Example 3-amino-1-(tbutyldiphenylsilyloxy)propane (470 mg, 1.5 minol) was acylated with tetradecanoyloxytetradecanoic acid (680 mg, 1.5 mniol) in the presence of EDC-MeI (595 mg, 2.0 inmol) and then deprotected with TBAF (1.0 M in THF, 2.0 mL, 2.0 inmol) in THF (10 mL) to afford 698 mg (91 of 3 3 -tetradecanoyloxytetradecanoylaminoy- 1 -propanol as an off-white solid.

In the same manner as described in Example the compound prepared in Example (7.9 g, 5.88 inmol) was deprotected with TFA (10 mL) and then treated with the Vilsmeier reagent generated from DMF (1.8 mL, 23.5 inmol) and WO 98/50399 WO 9850399PCT/US98/09385 64 oxalyl chloride (1.03 mL, 11.76 mmol) in CH 2

CI

2 (60 mL) to give 6.32 g (85 of 2deoxy-4-O-diphenylphosphono-3 -tetradecanoyloxytetradecanoyl60(2,22.

trichloro- 1, 1 -dimethylethoxycarbonyl)-2-(2,2,2trichloroethoxycarbonylamino)aDglucopyranosyl chloride as a white foam: 'H NMR (CDCI 3 8 0.8 8 6 H, J 6.8 Hz), 1.2 1.55 (in, 42 1.78 3 1.88 3 2.18 (t,2 H, J= 7.5 Hz), 2.43 (mn, 2 H), 4.31 (in, 4 4.68 1 H, J 11.9 Hz), 4.74 1 H, J= 11.9 Hz), 4.83 1 J= 9.3 Hz), 5.09 (in, 1 5.51 1 H, J 9.7 Hz), 5.78 1 H, J =8.0 Hz), 6.26 1 H, J= 3.4 Hz), 7.3 1 (in, 10 H).

In the same manner as described in Example 1 the compound prepared in above (613 ing, 1.2 mmol) and the compound prepared in above g, 1.2 inmol) were coupled in the presence of AgOTf (642 mng, 2.5 mmol) to give 1.43 g (68 of 3 3 -tetradecanoyloxytetradecanoylamino]propy 2-deoxy-4-0- [(R)-3-tetradecanoyoxytetradecanoyl]-60o(2,2,2-trichloro 1,1 dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-..D-glucopyranoside as an amnorphous solid: 'H NMR (CDC1 3 60.88 12 H, J=6.9 Hz), 1.1 -1.8(in, 86 1.82 3H), 1.89 3H), 2.20 2H, J=7.6 Hz), 2.29 2H,J= 7.7 Hz), 2.44 (mn, 4 3.21 (in, I 3.42 (mn, 1 3.54 (mn, 2 3.80 (mn, 1 3.94 (in, 1 4.28 (dd, 1 H, J= 12.3, 5.2 Hz), 4.3 8(d, 1 H, J= 10. 8Hz), 4.70 3H), 4.81 1 H, J =8.2 Hz), 5.14 (in, 2 5.47 I H, J 9.6 Hz), 6.13 I H, J =7.6 Hz), 6.22 (br. s, 1 H), 7.2 5 In the same manner as described in Example the compound prepared in above (700 ing, 0.40 iniol) was deprotected with zinc (1.32 g, 20.1 inmol) and then acylated with (R)-3-tetradecanoyloxytetradecanoic acid (200 ing, 0.44 inmol) in the presence of EEDQ (125 mg, 0.5 inmol) to give 435 mng (60 of tetradecanoyloxytetradecanoylaminolpropyI 2-deoxy-4-0-diphenylphosphono-3--[(R).

3-tetradecanoyoxytetradecanoyl]-2-[(R)-3 -tetradecanoyloxytetradecanoylamino])-p-Dglucopyranoside as an amorphous solid.

In the same manner as described in Example the compound prepared in above (400 mg, 0.22 inmol) was hydrogenated in the presence of platinum oxide (200 mg) to give 170 mg (45 of tetradecanoyloxytetradecanoylamino]propyl 2-deoxy-4-0-phosphono-3-0- WO 98/50399 WO 9850399PCTIUS98/09385 tetradecanoyoxytetradecanoyl]y2 -tetradecanoyloxytetradecanoylamino])43.D.

glucopyranoside triethylammonium salt as a white solid: mp 171-1720 C; JR (film) 3288, 3094, 2955, 2919, 2850, 1731, 1658, 1344, 1556, 1468, 1378, 1320, 1251, 1226, 11 72 ,110 6 ,10 8 3,1044 1H NMR (CDC 3

CD

3 OD) 6 0.88 18 H, J= 6.0 Hz), 1.1 1.7 (in, 135 2.2 -2.7 (in, 15 3.06 6 H, J= 7.1 Hz), 3.2 -4.1 (in, 8 4.21 1 H, J1=9.9 Hz), 4.51 I H, J= 8.3 Hz), 5.05 5.25 (mn, 4 7.23 1 H, J= 5.3 Hz), 7.33 1 H, J =8.6 Hz); 1 3 C NMR (CDCl 3 6 173.5, 173.4, 170.6, 170.2, 169.9, 100.6, 75.8, 71.5, 70.9, 70.5, 66.8, 60.4, 55.3, 45.6, 41.4, 39.4, 36.3, 34.6, 34.5, 34.2, 31.9, 29.7, 29.4, 29.3, 29.1, 25.4, 25.1, 22.7, 14.1, Anal. Calcd. for C 99

H

192

N

3 0, 7 P -2 C, 67.42; H, 11.20; N, 2.38; P, 1.76.

Found: C, 66.97; H, 11.01; N, 2.38; P, 1.95.

EXAMPLE 23 (B322) Preparation of 4 3 -Tetradecanoyloxytetradecanoylamino]butyl 2-Deoxy-4-Ophosphono 3 -3 tetradecanoyoxytetradecanoy ]2 tetradecanoyloxytetradecanoylamino])-..D-glucopyranoside Triethylammonium Salt (Compound

R,=R

2

=R

3 n-C1 3

H,

7 C0, X=Y=O, m=p=q=0O, R 4

==R

5

=R

6

=R

7

=R

9 q=H,

R

8 =P0 3

H

2 In the same manner as described in Example 20-(l1), 4-amino-i butyldiphenylsilyloxy)butane (500 mng, 1.53 mmol) was acylated with tetradecanoyloxytetradecanoic acid (695 mg, 1.53 mmol) in the presence of EDC-MeI (595 mng, 2.0 inmol) and then deprotected with TBAF (1.0 M in THF, 2.5 mL, 2.5 inmol) in THF (15 mL) to afford 651 mng (81 of tetradecanoyloxytetradecanoylamino]. A -butanol as an off-white solid.

In the same manner as described in Example 1 the compound prepared in above (650 mng, 1.25 mmol) and the compound prepared in Example 22- (1.6 g, 1.25 inmol) were coupled in the presence of AgOTf (L .16 g, 4.5 mmol) to give 1.65 g (75 of 4 3 -tetradecanoyloxytetradecanoylainino]butyl 2-deoxy-4-Odiphenylphosphono- 3 -O.[(R).3-tetradecanoyoxytetradecanoyl]6(22,2-trchr 1, 1 diinethylethoxycarbonyl)-2-(2,2,2..trichloroethoxycarbonylamino)-fo-D-glucopyranoside as an amorphous solid: 'H NMR (CDCl 3 6 0.88 12 H, J =6.9 Hz), 1. 1 1.8 (in, 88 1.82 3 1.89 3 2.15 2.55 (mn, 8 3.24 (in, 2 3.50 (in, 2 3.83 WO 98/50399 WO 9850399PCTIUS98/09385 66 (i,2 4.27 (dd, 1 H, J= 12.1, 3.8 Hz), 4.32 1 H, J= 11.5 Hz), 4.66 (in, 2 4.78 J H, J= 12.1 Hz), 4.89 I H,J =8.0Hz), 5.15 2H), 5.54 1 H, J =9.7 Hz), 5.95 (mn, 2 7.25 (mn, 10 H).

In the same manner as described in Example the compound prepared in above (700 mg, 0.39 minol) was deprotected with zinc (1.30 g, 19.8 mmol) and then acylated with 3 -tetradecanoyloxytetradecanoic acid (195 ing, 0.43 mmol) in the presence of EEDQ (125 mg, 0. 5 inmol) to give 421 mng (60 of 4- tetradecanoyloxytetradecanoylaminojbutyl 2-deoxy-4-O-diphenylphosphono-3...O[(R)-3tetradecanoyoxytetradecanoyl]-2- -tetradecanoyloxytetradecanoylamino])-3..D.

glucopyranoside as an amorphous solid.

In the same manner as described in Example the compound prepared in above (400 ing, 0.22 minol) was hydrogenated in the presence of platinum oxide (200 mg) to give 212 mng (55 of tetradecanoyloxytetradecanoylamino] butyl 2 -deoxy-4-O-phosphono-3 tetradecanoyoxytetradecanoyl] -tetradecanoyloxytetradecanoylainino])-3Dglucopyranoside triethylammoniuin salt as a white solid: mp 171-1720 C; IR (film) 3298,2955,2920,2851,1732,1645,1550, 1467, 1378,1181,1107, 1083, 1044,721 cn- I NMR (CDC1 3

-CD

3 OD) 8 0.88 18 H,J= 6.9 Hz), 1. 1 1.7 (in, 135 2.2 2.7 (in, 1 9 3.05 6H, J=7.1 Hz), 3.18 2H), 3.3 -3.5 6H)378(m, 3H),3.97 I H, J= 12.5 Hz), 4.23 I H, J= 10.0 Hz), 4.50 1 H, J= 8.5 Hz), 5.13 (in, 4 H), 7.12 1 H, J= 9.1 Hz); 1 3 CNMR (CDCl 3 8 173.9, 173.4, 173.3, 170.8, 169.9, 169.8, 101.0, 75.6, 73.2, 71.4, 71.1, 70.6, 68.9, 60.7, 54.8, 45.9, 41.5, 39.6, 38.9, 34.6, 34.3, 32.0, 29.8, 29.5, 29.0, 28.9, 26.3, 25.4, 25.1, 22.7, 14.2, 8.7.

Anal. Calcd. for C 100

H

194

N

3 0 17 P -H 2 0: C, 68.26; H, 11.23; N, 2.39; P, 1.76.

Found: C, 68.21; H, 11.03; N, 2.26; P, 1.73.

WO 98/50399 PCT[US98/09385 67 EXAMPLE 24 (B23) Preparation of 4- -Tetradecanoyloxytetradecanoylamino]hexyI 2-Deoxy-4-Op h os ph on o-3 0- -3 t etr ad e canoy o xyt e trad ec an o y I -2 tetradecanoyloxytetradecanoylamino].p-D-glucopyranoside Triethylammonium Salt (Compound R,=R 2

=R

3 n-C 3

H

27 C0, X=Y=O, n=4, m=p=q=0, R 4

=R

5

=R

6

=R

7

=R

9

=H,

R

8 =p0 3

H

2 In the same manmer as described in Example 20-( 6-amino-I butyldiphenylsilyloxy)hexane (1.48 g, 4.15 mmol) was acylated with tetradecainoyloxytetradecanoic acid (2.07g, 4.56 mmol) in the presence of EDC-MeI (1.35g, 4.56 mmol) and then deprotected with TBAF (1.0 M in THF, 1.53 mL, 1.53 mmol) in THF (46 mL) to afford 700 mg (30 of tetradecanoyloxytetradecanoylamino]- 1 -hexanol as an off-white solid.

In the same manner as described in Example the compound prepared in above (689 mg, 1.20 mmol) and the compound prepared in Example 22- (1.25 g, 1.00 mmol) were coupled in the presence of AgOTf (1.28 g, 5.0 mmol) to give 1.59 g (94 of 4 -[(R)-3-tetradecanoyloxytetradecanoylamino]hexyl 2-deoxy-4-0diphenylphosphono-3-0-[(R)-3 -tetradecanoyoxytetradecanoyl]-6-0-(2,2,2..trichloro- 1, 1 dimethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylamino)-pD.glUCopyranoside as an amorphous solid: 'H NMR (CDCl 3 60.88 12 H,J= 6.6 Hz), 1.1 -1.8 (in,92 1. 82 3 1. 89 3 2.22 2 H, J =7.6 Hz), 2.29 2 H, J1 7.4 Hz), 2.45 (in, 4 3.22 (in, 1 3.46 (in, 2 3.83 (in, 2 3.94 (in, I 4.31 (mn, 2 4.64 (in, 2 4.83 1 H, J 12.1 Hz), 4.97 I H, J 7.8 Hz), 5.17 (in, 2 5.59 1 H, J 8.8 Hz), 5.7 5 (in, 1 5.84 1 H, J 7.6 Hz), 7.25 (in, 10 H).

In the same manner as described in Example the compound prepared in above (1.57 g, 0.88 inmol) was deprotected with zinc (2.88 g, 44.1 mrnol) and then acylated with (R)-3-tetradecanoyloxytetradecanoic acid (481 mng, 1.06 immol) in the presence of EEDQ (327 mng, 1.32 inmol) to give 1.57 g (97 of tetradecanoyloxytetradecanoylamino]hexyl 2-deoxy-4-0-diphenylphosphono-3-0-[(R)-3tetradecanoyoxytetradecanoyl]-2- -tetradecanoyloxytetradecanoylamino])-3-D.

glucopyranoside as an amorphous solid.

In the same manner as described in Example the compound prepared in above (1.57 g, 0.85 minol) was hydrogenated in the presence of platinum WO 98/50399 WO 9850399PCT/US98/09385 68 oxide (157 mg) to give 130 mg (10 of tet;adecanoyloxytetradecanoylamino]hexyl 2-deoxy-4-O-phosphono..3..O[(R-3 tetradecanoyoxytetradecanoyl] -tetradecanoyloxytetradecanoylamino] -j-Dglucopyranoside triethylammoniumn salt as a white solid: mp 150-152' C; IR (film) 3284, 3099, 2954, 2920, 2851, 1731, 1657, 1637, 1557,1467, 1418, 1378, 1320, 1249, 1 1 7 9 1 1 0 8 ,1 0 8 3 ,10 4 4 8 56,721lcm-'; 'H NMR(CDCI 3

-CD

3 OD) 80.89 18 H, J= 6.6 Hz), 1. 1 1.7 (in, 13 5 2.2 -2.7 (mn, 23 3.05 6 H, J= 7.1 Hz), 3.18 (in, 2 3.39 1 H, J 8.2 Hz), 3.49 1 H, J 7.5 Hz), 3.82 (mn, 2 3.99 1 H, J 11.9 Hz), 4.25 1 H, J 8.9 Hz), 4.59 (mn, 2 5.18 (in, 4 3 C NMR (CDCl 3 6 173.7, 173.3, 170.6, 169.7, 169.4, 100.6, 75.5, 73.1, 71.3, 70.9, 70.6, 69.2, 60.6, 55.2, 45.8,41.7,41.4,39.5,39.4,34.6,34.3,34.2,34.1,31.9,29.7,29.4, 29.2,26.5,25.5,25.3, 25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for CJQ 2 H,1 98

N

3 01 7 P, H 2 0: C, 68.53; H, 11.28; N, 2.33; P, 1.73.

Found: C, 68.63; H, 11. 12; N, 2.26; P, 1.66.

EXAMPLE 25 (B324) Preparation of -Tetradecanoyloxytetradecanoyl] -O-phosphono-2- tetradecanoyloxytetradecanoylamino]-3-O..[(R)-3-tetradecanoyloxytetradecanoyl]-..D.

glucopyranosyl]-L-serinainide Triethylainmoniuin Salt (Compound R,=R 2

=R

3 =n- C, 3

H

27 C0, X=Y=O, n=m=p=q=0, R 4

=R

5

=R

7

=R

9 R6=CONH 2

R

8 =P0 3

H

2 A suspension of L-serinamide hydrochloride 15 7 g, 1. 18 mmol) and (R)-3-tetradecanoyloxytetradecanoic acid (0.61 g, 1.34 mmol) in CH 2 Cl 2 (6 mL) was treated with triethylamine 18 mL, 1. 3 inmol) and the resulting solution was stirred with 4 A molecular sieves for 30 min. EEDQ (0.43 7 g, 1.77 minol) was then added and the mixture was stirred for 16 h at room temperature. The product that precipitated was collected and washed with CH 2 Cl 2 (2 x 25 mL) to give 0.455 g (7 of tetradecanoyloxytetradecanoyl]-L-serinamide as a colorless powder: mp 126-130'C; 'H NMR (CDCI 3 860.88 6H, J= ~7 Hz), 1. 15-1.7 (in, 42 2.31 2 H, J=7.5 Hz), 2.51 2 H, 1=6.3 Hz), 3.56 (br s, 1 3.65 (dd, I H, J=1 1.2, 5.5 Hz), 3.86 (dd, 1 H, J =1 1.2, 4.5 Hz), 4.21 2 4.40 (in, 1 5.22 (in, 1 H).

In the same manner as described in Example the compound prepared in above (0.23 g, 0.246 inmol) and the compound prepared in Example 2-(4) WO 98/50399 WO 9850399PCT[US98/09385 69 (0.961 g, 0.745 mmol) were coupled in the presence of mercury cyanide (0.43 g, 1.7 mmaol) to give 0.527 g (71 ofN-[(R)- 3 -tetradecanoyoxytetradecanoyyo-[2-deoxy-4 0-diphenylphosphono-3-0- 3 -tetradecanoyloxytetradecanoy16.o(22,2trichloro- 1, 1 -dimethylethoxycarbony1)-2-(2,2,2,trichloroethoxycarbonylamjflo)f-DglucopyranoSyl]-L-serinamide as an amorphous solid: 'H NMR (CDCl 3 6 0.88 12 H, J= -7 1.0-1.7 (in, 84 1.80 and 1.89 (2s, 6 2.21 2 H, J=7.5 Hz), 2.30 2 H, J=7.5 Hz), 2.37 (in, 2 2.47 (mn, 2 3.54 (mn, 1 3.68 (dd, 1 H, J=8, J=l1 IHz), 3.86 (br d, 1 H, J=1 1 Hz), 4.16 (dd, 1 H, J=1 1, 4Hz), 4.24 (dd, I H, J=12, 4.3 Hz), 4.40 1 H, J= 12 Hz), 4.6-4.8 (mn, 4 5.00 1 H, J=8 Hz), 5.1-5.25 (in, 2 5.4-5.55 (in, 2 5.84 (br s, 1 6.61 (br s, 2 7.1-7.3 5 (in, 10 H).

In the same manner as described in Example the compound prepared in above (0.44 g, 0.254 minol) was deprotected with zinc (0.83 g, 13 minol) and then acylated with (R)-3-tetradecanoyloxytetradecanoic acid 14 g, 0.31 mmnol) in the presence of EEDQ (0.095 g, 0.38 inmol) to give 0.27 1 g of tetradecanoyloxytetradecanoyl] -0-[2-deoxy-4- 0-diphenylphosphono-2- tetradecanoyloxytetradecanoylamino-3...o[(R)-3 -tetradecanoyloxytetradecanoyl]-f3Dglucopyranosyl]-L-serinamide as an amorphous solid: 'H NMR (CDCI 3 6 0.88 18 H, Hz), 1.0-1.7 (in, 126 2.03 (br s, 1 2.15-2.55 (in, 12 3.5-4.05 (mn, 5 H), 4.14 (dd, 1 H, J= 10, 3.5 Hz), 4.5-4.65 (in, 2 4.68 1 H, J=8.1 Hz), 5.05 -5.25 (in, 3 5.31 1 H,J= 10 Hz), 5.5 8 (br s, 1 6.31 1 H, J=8 Hz), 6.85-6.95 (in, 2 7.1-7.4 (in, 10 H).

In the same manner as described in Example the compound prepared in above (0.25 g, 0. 14 inmol) was hydrogenated in the presence of platinum oxide (0.125 g) to give 0.195 of N-[(R)-3-tetradecanoyloxytetradecanoyl].o.[2.

deoxy-4-0-phosphono-2- -tetradecanoyloxytetradecanoylamino] tetradecanoyloxytetradecanoyl-D-glucopyranosyl]yL-serinamide triethylainmoniumn salt as a colorless solid: mp 190-191 'C (dec); IR (film) 3418, 3293, 2921, 2850, 1732, 1717, 1651, 1636, 1557, 1540, 1458, 1165, 1033 'HNMR (CDCI 3

-CD

3 OD) 80.88 18 H, J= 7Hz), 1.0- 1.7 (in, 13 5 2.2-2.7 (in, 12 3.05 6 H, J=7.2 Hz), 3 .2- 3.45 3.5-4.15 (mn,5 4.21 I H, 10OHz), 4.53 1 H, J=8.1 Hz), 4.58(mn, I 5.0-5.3 (in, 4 7.25 1 H, J=8.4 Hz), 7.40 1 H, J=7.2 Hz); 1 3 C NMR I. WO 98/50399 WO 9850399PCTIUS98/09385 (CDCl 3

-CD

3 OD) 8 173.7, 173.5, 172.5, 170.7, 170.5, 170.4, 101.4, 75.5, 73.4, 71.1, 70.9, 70.2,68.6,60.0, 53.9, 52.2,45.6,41.2,41.0, 38.9, 34.4, 34.2,31.8,29.6,29.5,29.3,29. 1, 25.2, 24.9, 22.6, 14.0, 8.3.

Anal. Calcd for C 99

H

191

N

4 0 18 P -2.5 H,O: C, 66.00; H, 10.97; N, 3.11; P, 1.72.

Found: C, 66.04; H, 10.99; N, 3.03; P, 1.95.

EXAMPLE 26 Preparation ofN- [(R)-3-Decanoyloxytetradecanoyl]-0- [2-deoxy-4-0-phosphono-2-[(R).

3 -decanoyloxytetradecanoylamino] -decanoyloxytetradecanoyl]-p-D.

glucopyranosyl]-L-serinamide Triethylammonium Salt (Compound R, =R 2

=R

3 =n-

C

9 H1 9 C0, X=Y=O, nm=p=q=0, R 4

=R

5

=R

7

=R

9 R6=CONH,,R 8

=PO

3

H

2 In the same manner as described in Example 25 L-serinamide hydrochloride (169 mg, 1.2 mmol) was acylated with (R)-3-decanoyloxytetradecanoic acid (478 mg, 1.2 mmol) in the presence of EEDQ (371 mg, 1.5 mniol) in CH 2 Cl, to afford 428 mg (74 of N-[(R)-3-decanoyloxytetradecanoyl]-L-serinamide as a white solid: 'H NMR (CDC1 3 6 0.88 6 1. 1 1.7 (in, 34 2.33 2 H, J 7.5 Hz), 2.54 2 H, J= 6.6 Hz), 3.35 2 3.72 (dd, I H, J= 11.0, 5.2 Hz), 3.84 (dd, I H, J 11. 3, 5.0 Hz), 4.20 1 H, J 5.1 Hz), 5.26 1 H, J 6.4 Hz).

In the same manner as described in Example the compound prepared in above (410 mg, 0.85 minol) and the compound prepared in Example (1.05g, 0.87 mmol) were coupled in the presence of AgOTf (560 mg, 2.2 mmol) to afford 780 g (56 of N-[(R)-3-decanoyloxytetradecanoyl]-0-[2-deoxy-4...odipheny phosphono-3-0-[(R)-3-decanoyloxytetracecanoyl60(2,2,2-trichloro. 1 1 diinethylethoxycarbony)-2-(2,2,2-trichloroethoxycarbonyanino)o.D-glucopyranosy].

L-serinainide as an amorphous solid: 'H NMR (CDCl 3 6 0. 88'(t, 12 1. 1 1.6 (in, 68 1.80 3 1.89 3 2.30 (in, 8 3.53 (in, 1 3.68 (in, 1 3.85 (br. d, 1 H, J= 9.4 Hz), 4.15 (dd, 1 H, J= 10.8, 3.7 Hz), 4.24 (dd, I H, J= 12.3, 4.6 Hz), 4.40 I H, J= 10.8), 4.65 (in, 4 5.00 I H, J= 8.2 Hz), 5.18 (in, 2 5.46 (in, 2 H), 5.83 (in, I 6.60 (in, 2 7.30 (in, 10 H).

In the same manner as described in Example the compound prepared in above (600 mng, 0.36 inmol) was deprotected with zinc (1.19 g, 18.2 mmol) and acylated with (R)-3-decanoyloxytetradecanoic acid 16 0 mg, 0.4 inmol) in the WO 98/50399 WO 9850399PCTIUS98/09385 71 presence of EEDQ (124 mg, 0.50 mmol) to afford 371 mg (62 of deranoyloxytetradecanoyl [2-deoxy-4- 0-diphenylphosphono-2- decanoylIo xytetradecanoyl amino] -3 0- -3 -decanoyloxytetradecanoylI] 0 -Dglucopyranosyl]-L-serinamide as an amorphous solid.

In the same manmer as described in Example the compound prepared in above (330 mg, 0.20 mmol) washydrogenated in the presence of platinum oxide (200 mg) to afford 120 mg (44 of N-[(R)-3-decanoyloxytetradecanoyl]pO.42-.

deoxy-4-0-phosphono-2- 3 -decanoyloxytetradecanoylIamino] 3 0- decanoyloxytetradecanoy1]-i-D-glucopyranosyl]Lserinmide triethylammonium salt as a white powder: mp 187-189' C; IR (film) 3419, 3286, 3220, 3 098, 2955, 2922, 2852, 1732, 1680, 1662, 1644, 1559, 1467, 1247, 1167, 1107, 1080, 1051, 965, 913 'H NMR (CDC 3

CD

3 OD) 860.89 18 H, J =7.0 Hz), 1. 1 1.7 (in, 111 2.2 -2.7 (in, 12 3.07 6 H, J= 7.1 Hz), 3.68 (in, 1 3.87 (in, 1 4.09 (dd, 1 H, J= 10.8, 3.6 Hz), 4.22 (in, 1 4.5 3 1 H, J1= 8.2 Hz), 4.5 8 (in, 1 5.13 (in, 3 5.2 8 (in, 1 H), 7.53 I H, J =9.0Hz), 7.56(d, 1 H,J =7.7 Hz); 1 3 CNMR (CDC1 3 8 173.5, 173.2, 170.2, 169.8, 102.3, 75.7, 73.5, 71.3, 70.7, 70.1, 68.8, 60.8, 53.9, 51.7, 45.8, 41.5, 4 1. 1, 39.1, 34.6, 34.5, 34.2, 32.0,29.7, 29.6, 29.5, 29.4, 25.7, 25.4, 25.1, 22.7, 14.1, 8.6.

Anal. Calcd. for C 87

H

167

N

4 0 18 P C, 65.05; H, 10.60; N, 3.49; P, 1.93.

Found: C, 65.06; H, 10.40; N, 3.3 1; P, 2.00.

EXAMPLE 27 (B26) Preparation ofN-[(R)-3-Tetradecanoyloxytetradecanoyl]-0-[2-deoxy..4-0-phosphono2- [(R)-3-tetradecanoyloxytetradecanoylainino]-3-0- [(R)-3-tetradecanoyloxytetradecanoyl]- P3-D-glucopyranosyl]-L-serine Methyl Ester Triethylanimoniun Salt (Compound

R,=R

2

=R

3 =n-C3 1

H

2 7 C0, X=Y=O,

R

4

=R

5

=R

7

=R

9 R6=CO 2 Me,

R

8 =P0 3

H

2 A solution of the compound prepared in Example 12-(2) (0.290 g, 0. 157 inmol) in THF (20 mL) was hydrogenated in the presence of 5% palladium on carbon mg) at room temperature and atmospheric pressure for 3 h. The catalyst was removed by filtration and the filtrate concentrated. A solution of the residue in CHC1 3 mL) at 0 0 C was treated with a solution of diazoinethane (0.5 minol) in ether (5 mL) and then stirred for 30 min at 0 0 C. AcOl- (0.5 inL) was added and the resulting colorless k, WO 98/50399 WO 9850399PCTIUS98/09385 72 solution was diluted with ether (50 mnL), washed with saturated aqueous NaHCO 3 mLU), dried (Na 2

SO

4 and concentrated. Flash chromatography on silica gel (gradient elution, 20 25% EtOAcs-hexanes) afforded 0.199 g of tetradecanoyloxytetradecanoyl]po [2-deoxy-4-O-diphenylphosphono-3-0- tetradecanoyloxytetradecanoyl]6.0-(2 ,2,2-trichoro- 1, 1 -dimethylethoxycarbonyl)2- 2 2 2 -trichloroethoxycarbonylamino)-p-D.glucopyrnosyl]-Lserine methyl ester as an amorphous solid: 'H NMR (CDCl 3 60. 88 12 H, J= 6.5 Hz), 1.1- 1.75 (mn, 84 1.81 and 1.89 (2s, 6 2.36 2 H, J=7.5 Hz), 2.25-2.6 (in, 6 3.48 1 H, J= -8 Hz), 3.7-3.9 (in, 5 4.2-4.4 (in, 3 4.6-4.85 (in, 4 4.88 1 H, J=7.8 Hz), 5.03-5.22 2H), 5.49 1 H, J= -9.5 Hz), 6.21 (br s, I 6.59 1 H, J=7.8 Hz), 7.1-7.4 (m,

H).

In the same manner as described in Example the compound prepared in above 195 g, 0. 111 mnmol) was deprotected with zinc 36 g, minol) and acylated with 3 -tetradecanoyloxytetradecanoic acid (0.060 g, 0. 13 inmol) in the presence of EEDQ (0.041 g, 0. 17 mmol) to give 0. 138 g of tetradecanoyloxytetradecanoyl] -4-0-diphenylphosphono.2- tetradecanoyloxytetradecanoyaino]3o0[(R-3-tetradecanoyloxytetradecanoyl-p-D glucopyranosyl]-L-serine methyl ester as an amorphous solid: 'H NMR (CDCl 3 )560.88 18 H, J= -6.5 Hz), 1.0- 1.75 (in, 126 2.15-2.45 (in, 10 2.52 (dd, 1 H, J=1 4.7, 6 Hz), 2.66 (dd, 1 H, J =14.7, 6 Hz), 3.3 5 (br s, I 3.4-3.8 (in, 7 3.8 8 (dd, I H, J =11I Hz), 4.18 (dd, I H, J=1 I Hz), 4.6-4.75 (mn, 2 5.03 1 H, J=7.8 Hz), 5.1-5.25 (in, 3 5.5 0 1 H, J= Hz), 6.5 0 1 H, J=7.2 Hz), 6.97 1 H, J=7.8 Hz), 7.1 7.4 (in, 10 H).

In the same manner as described in Example the compound prepared in above (0.100 g, 0.055 minol) was hydrogenated in the presence of platinum oxide (50 ing) to give 0.055 g of tetradecanoyloxytetradecanoyl] -0-[2-deoxy-4-O-phosphono.2.[(R)-3 tetradecanoyloxytetradecanoylainino-3 3 -tetradecanoyloxytetradecanoyl]-o-.Dglucopyranosyl]-L-serine methyl ester triethylainmoniuin salt as a colorless solid: mp 142-143TC(dec); IR(film) 3289,2955,2921, 2852,1733,1718,1699,1652,1558,1540, 1521, 1506,1469,1457,1375,1360, l259cm-'; 'HNMR(CDC 3

-CD

3 OD)80.88 18 WO 98/50399 PCTIUS98/09385 7 3 H, J= 6.5 Hz), 1.0- 1.7 (in, 13 5 2.2-2.7 (mn, 12 3.05 6 H, J=7.5 Hz), 3.31 (d, 1 HK J=9.3 Hz), 3.37 I 3.55-3.9 (mn, 10 3.97 1 H, J=1 2 Hz), 4.1-4.25 (mn, 2 4.55-4.65 (mn, 2 5.05-5.25 (in, 3 7.23 1 H, .1=8.1 Hz), 7.47 1 H, .1=7.2 Hz); 1 3

CNMR(CDC

3 6 173.6, 173).4, 170.5, 170.4, 170.1, 100.7,75.9,72.8,71.2, 70.8, 70.6,68.5,60.3,55.3,52.7, 52.4,47.7,41.5,40.9, 39.7,34.6,34.5,34.3. 32.0,29.8,29.4, 25.4, 25.1, 22.7, 14.2, Anal. Calcd for C 100

H

192

N

3 01 9 P -H 2 0: C, 67.11; H, 10.93; N, 2.35; P, 1.73.

Found: C, 66.9 1; H, 10.93; N, 2.3 1; P, 2.11.

EXAMPLE 28 (B27) Preparation of N-(Carboxymethyl)-N-[(R)-3 -tetradecanoyloxytetradecanoyl..2aminoethyl 2 -Deoxy- 4 -O-phophono-2-[(R)-3-tetradecanoyloxytetradecanoylamino].3 O-[(R)-3-tetradecanoyloxytetradecanoyl] -f3--glucopyranoside Triethylammoniumn Salt (Compound R 1

=R

2

=R

3 =n-C 1 3

H

27 C0, X=Y=O, n=m=p=0, R 4

=R

5 =R6=Rq=H,

R

7

=CO

2 H, q=1, Rs=PO 3 In the same manner as described in Example N-(2hydroxyethyl)glycine t-butyl ester (0.25 g, 1.43 rmol) was acylated with tetradecanoyloxytetradecanoic acid (0.714 g, 1.57 minol) in the presence of EDC-MeI (0.466 g, 1.57 mmol) to give 0.46 g (5 of N-(2-hydroxyethyl)-N-[(R)-3tetradecanoyloxytetradecanoyllglycine t-butyl ester as an amorphous solid: 1H NMR (CDCl 3 60.88 6 H, 6.5 Hz), 1.15-1.7 51 2.26 2H, J =7.5 Hz), 2.60 (dd, I H, J 6.5, 15 Hz), 2.86 (dd, 1 H, J 15 Hz), 3.40-4.15 (in, 7 5.25 In1 H).

In the same mannner as described in the compound prepared in above (0.21 g, 0.334 iniol) and the compound prepared in Example 22-(2) (0.458g, 0.368 inmol) were coupled in the presence of AgOTf (0.688 g, 2.68 inmol) to give 0.39 g of N-(t-butyloxycarbonylnethyl)-N-[(R)-3tetradecanoyloxytetradecanoyl]-2-aminoethyl 2 -deoxy-4-O-diphenylphosphono-3 -0- -tetradecanoyloxytetradecanoylI-6-0-(2,2,2-trichloro 1,1 diinethylethoxycarbonyl)-2-(2,2,2-trichloroethoxycarbonylainino)-..Dglucopyranoside as an amorphous solid: 'H NMR (CDCI 3 60.88 (t,12 H, Hz), 1.0- 1.95 (in, 99 2.1-2.6 (in, 7 2.84 (dd, 1 H, J= 5, 15 Hz), 3.2-4.15 (in, 8 WO 98/50399 WO 9850399PCTIUS98/09385 74 4.15-4.45 (in, 2 4.55-4.9 (in, 3 5.00 1 H, J1= 8 Hz), 5.13 (in, 2 5.4- 5.65 (mn, 1 6.16 I H, J 7 Hz), 7.05-7.4 (in, 10 H).

In the same manner as described in Example the compound prepared in above (0.339g, 0. 185 mmol) was deprotected with zinc (0.36 g, 5.54 inmol) and then acylated with 3 -tetradecanoyloxytetradecanoic acid 100 g, 0.221 mmol) in the presence of EEDQ (0.068 g, 0.276 inmol) to give 0.25 g of N-(t-butyloxycarbonylmethyl).N-.[(R)-3 -tetradecanoyloxytetradecanoyl]y2.aminoethyI 2-deoxy-4-O-phosphono-2- -tetradecanoyloxytetradecanoylamino-3 tetradecanoyloxytetradecanoyl]--D-glucopyranoside as a colorless solid.

In the same manner as described in Example the compound prepared in above (0.25 g, 0. 131 mmol) was hydrogenated in the presence of platinum oxide (125 mg) in 9:1 THF-AcOH (15 mnL). The crude hydrogenolysis product was dissolved in CH 2 C1 (1 mL), cooled to 0 0 C, and treated dropwise with TFA (0.5 inL). After stirring for 2 h at 0 0 C, the reaction mixture was concentrated and residual TFA was removed by azeotroping with toluene. The resulting residue (0.23 g) was dissolved in 1% aqueous triethylainine (12 mL) and lyophilized. Flash chromatography on silica gel with chlorofornmethanol-water-triethylamine (91:8:0.5:0.5-.85:15:0.5:0.5, gradient elution) and further purification by means of acidic extraction as described in Example and lyophilization from 1% aqueous triethylamnine (6 mL) afforded 99 mng of N-(carboxymethyl)-N-[(R).3.

tetradecanoyloxytetradecanoyl].2-aminoethy 2-deoxy-4-0-phosphono-2[(R)-3 tetradecanoyloxytetradecanoylanino]-3 -tetradecanoyloxytetradecanoyl]-3- D-glucopyranoside triethylammonium salt as colorless solid: mp 162-163'C (dec); IR (film) 3286, 2922, 2852, 1732, 1651, 1556, 1455, 1434, 1378, 1260, 1088, 801 cm-r 'H NMR (CDCl 3 60.88 18H, Hz), 1.0-1.75 (in,13S 2.2-3.0 14 3.04 6 H, J 7.2 Hz), 3.2 5-3.8 (mn, 5 3.8 5-4.3 (in, 5 4.5 5 1 H, J= Hz), 4.68 1 H, J= 8.1 Hz), 5.05-5.35 (in, 4 H).

Anal. Calcd for C, 00

H

192

N

3 0 19 P 3 H 2 0: C, 65.79; H, 10.60; N, 2.30; P, 1.70. Found: C, 65.82; H, 10.44; N, 2.40; P, 1.79.

0 WO 98/50399 PCTIUS98/09385 EXAMPLE 29 (1328) Preparation of N-Carboxymethyl-N[(R)3decanoyloxytetradecanoyl3aminopropyl 2 -Deoxy- 4 -O.phosphono-2[(R)3decanoyloxytetradecanoylamino)-3 decanoyoxytetradecanoyl]-f3-n-glucopyranoside Triethylammonium Salt (Compound

R

1

=R

2

=R

3 n-C 9

H,

9 C0, X=Y=O, n 1, mp=0, R 4

=R

5

=R

6

=R

9 R7=CO 2 H, q=1,

R

8 =P0 3

H

2 In the same manner as described in Example hydroxypropyl)glycine, benzyl ester (450 mg, 2.0 mmol) was acylated with decanoyloxytetradecanoic acid (1.0 g, 2.5 mmol) in the presence of EDC-MeI (900 mg, 3.0 mmol) in CH 2 C1, to afford 0.76 g (63 of N-(3-hydroxypropyl)-N-[(R)-3-.

decanoyloxytetradecanoyljglycine benzy] ester as a colorless oil: 'H NMR (CDCl 3 (1:1I mixture of rotomers) 0.88 6HJ =6.6 Hz), 1. 1- 1.7 35 1.78 1 2.26 2 H, J 7.6 Hz), 2.3 7 and 2.54 (2 dd, 1 H, J =14.9, 6.9 Hz), 2.60 and 2.89 (2 dd, I H, J 14.8, 6.0 Hz), 3.51 (in, 4 3.70 (mn, 1 3.95 4.25 (in, 2 H), 5.1 5.25 (in, 3 7.35 (in, 5 H).

In the same manmer as described in Example the compound prepared in above (500 ing, 0.83 minol), and the compound prepared in Example 15-(4) (1.0g, 0.83 inmol) were coupled in the presence of AgOTf (1 .07 g, 4.15 mmol) to afford 1.27 g (72 of N-(benzyloxycarbonylmethyl)-N[(R)-3 decanoyloxytetradecanoyl]-3-aminopropyl 2-deoxy-4-O-diphenylphosphono.3..0- -decanoyoxytetradecanoyl3-6-o-(2,2,2-trichloro. ,1 -di methylethoxycarbonyl)- 2 2 2 2 -trichloroethoxycarbonylamino)-3-D-glucopyranoside benzyl ester: 'H NMR (CDCl 3 (2 1 mixture of rotomers) 8 0.88 12 H, J= 6.9 Hz), 1.1 1.7 (in, 69 H), 1.80 3H), 1.88 3H), 2.1 2.6 I11 2.81 (dd, I H, J= 14.8, 6.2 Hz), 3.37 (in, I 3.52 (in, 2 3.76 (in, 1 3.87 (in, 1 4.05 (in, 2 4.28 (in, 3 H), 4.62 (in, 3 4.77 (in, 1 4.93 1 H, J= 8.2 Hz), 5.15 (in, 4 5.46 and 5.61 (2 t, I H, J =9.5Hz),5.95 and6.05 (2d, 1 H, J =7.5Hz), 7.1 7.4 15 H).

In the same manner as described in Example the compound prepared in above (1.25 g, 0.71 inmol) was deprotected with zinc (2.31 g, 3.53 inmol) and acylated. with 3 -decanoyloxytetradecanoic acid (353 ing, 0.89 inmol) in the presence of EEDQ (264 ing, 1.07 nunol) to afford 670 mng (54 of Nbenzyloxycarbonylmethyl-N-[(R-3 -decanoyloxytetradecanoyl]-3-aminopropyI 2- WO 98/50399 PCT/US98/09385 76 deoxy-4-O-diphenylphosphono-3-0-[(R)-3-decanoyoxytetradecanoyl]-2-[(R)-3desanoyloxytetradecanoylamino])-3-D-glucopyranoside as an amorphous solid.

In the same manner as described in Example the compound prepared in above (670 mg, 0.38 mmol) was hydrogenated in the presence of palladium hydroxide on carbon (270 mg) and platinum oxide (200 mg) in EtOH AcOH (10:1) to afford 240 mg (39 of N-carboxymethyl-N-[(R)-3decanoyloxytetradecanoyl]-3-aminopropyl 2-deoxy-4-0-phosphono-2-[(R)-3decanoyloxytetradecanoylamino])-3-O-[(R)-3-decanoyoxytetradecanoyl]-P-Dglucopyranoside triethylammonium salt as a white powder: mp 156 1570 C; IR (film) 3284, 2929, 2853, 2729, 1732, 1655, 1628, 1551, 1466, 1378, 1314, 1164, 1108, 1047, 955, 844, 722 'H NMR (CDC 3 CD30D) 6 0.88 18 H, J= 6.9 Hz), 1.1 1.7 111 2.27 6 H, J= 6.2 Hz), 2.35 2.80 9 3.05 6 H, J= 7.2 Hz), 3.25 3.60 4 3.75 4.10 4 4.23 2 4.47 1 H, J= 8.2 Hz), 4.61 1 H, J= 8.3 Hz), 5.05 5.25 4 3 C NMR (CDC13) 8 173.4, 173.0, 171.1, 170.6, 170.3, 169.6, 100.5, 74.5, 73.9, 71.4, 71.2, 70.7, 70.2, 67.0, 65.8, 60.7, 54.6, 54.3, 51.4, 49.2, 46.0, 45.4, 42.1, 41.2, 39.4, 38.0, 37.7, 34.5, 34.3, 34.2, 31.9, 29.8, 29.7, 29.6, 29.5, 29.2, 28.1, 25.4, 25.3, 25.1, 22.7, 14.1, 11.1, 8.6.

Anal. Calcd. for C 89

H,

70

N

3 0, 9 P H 2 0: C, 65.37; H, 10.60; N, 2.57; P, 1.89.

Found: C, 65.35; H, 10.42; N, 2.43; P, 2.05.

TEST EXAMPLE 1 Stimulation of Anti-tetanus Toxoid Antibody Production.

The AGPs of the subject invention enhanced antibody production to purified tetanus toxoid in a murine model. Ten mg of each AGP sample was added to 1 ml of an oil-lecithin mixture containing squalene oil plus 12% lecithin. The mixtures were heated in a 56 °C water bath and sonicated to achieve clear solutions. Fifty (50) Al of each solution was emulsified by vortexing in 2 ml of sterile, pre-warmed 0.1% Tween saline containing 1.0 .g tetanus toxoid antigen/ml. Preparations were vortexed again just prior to administration to mice. Female C57BL/6 x DBA/2 F, mice (8 per group) were treated with 0.2 ml of the appropriate preparation distributed as a 0.1 ml WO 98/50399 PCT/US98/09385 77 subcutaneous injection into each flank. The final mouse dosage of the tetanus toxoid and AGP compounds was 0.2 lg and 50 1g, respectively. Control mice received tetanus toxoid in vehicle (oil-Tween saline). All mice were treated on day 0 followed by a second immunization on day 21. Fourteen days following the second immunization mice were bled and sera were isolated by centrifugation.

Serum samples from each mouse were evaluated for anti-tetanus toxoid antibodies by enzyme immunoassay (EIA) analysis using tetanus toxoid coated microtiter plates. Anti-tetanus antibody titers were evaluated for IgM, total Ig, as well as, IgG,, IgG2a and IgG2b isotypes. Each serum sample was diluted 2-fold for eleven dilutions starting with an initial serum dilution of 1:200. Results are shown in Tables 2-4.

0 Table 2.

Anti-tetanus toxoid antibody titers of treated mice.

Material B311 B2 B I B21 Total IgG T/C* Titer 3.6 23,200 3.84 24,800 3.97 25,600 8.93 57,600 4.71 30,400 18.85 121,600 6,450 IgG 1 TIC Titer 1.86 400,000 2.16 464,000 3.42 736,000 2.68 576,000 2.23 480,000 4.17 896,000 215,000

TIC

2.06 4.28 3.78 1.67 5.83 6.80 Titer 10,450 21,700 19,200 8,500 29,600 34,500 5,075

TIC

0.93 1.57 IgG 2 IgG 2 b Titer 26,800 45,200

TIC

4.75 4.50 2.38 2.0 1gM Titer 7,600 7,200 3,800 3,200 2.45 70,400 3.28 94,400 6.07 174,400 2.79 80,256 28,750 5.50 8,800 6,400 1,600 Vehicle *T/C Ratio=Experimental Test Titer Vehicle Control Titer.

WO 98/50399 WO 9850399PCTIUS98/09385 R19702CI Table 3.

Anti-tetanus toxoid antibody titers of treated mice.

Material B 12 B16 B13 BlI Vehicle

TIC*

3.1 1.6 3.9 3.3 1gM 4800 2560 6080 5120 1760

TIC

139.4 66.8 220 347 IgG 2a 2370 1135 3740 5900 25

TIC

149 104 >208 127.3 IgG 2 b 9840 6880 13,760 8400 98 *T/C Ratio= Experimental Test Titers -Vehicle Control Titers I Table 4. 0 Anti-tetanus toxoid antibody titers of treated mice.

Material Total Ig IgM IgG, IgG 2 IgG2 b T/C Titer T/C Titer T/C Titer T/C Titer T/C Titer B26 10.5 2,490 1.1 600 16.9 25,200 29.3 440 42.6 2,260 144.5 34,400 2.7 1,520 118.3 176,000 259.3 3,890 603.8 32,000 B22 60.0 19,050 0.8 440 18.4 27,400 345.8 5,187 59.6 3,160 B28 228.6 54,500 3.7 2,080 92.5 137,600 664.7 9,970 519.2 27,520 Vehicle 238 560 1,488 15 53 *T/C Ratio=Experimental Test Titer Vehicle Control Titer.

I

WO 98/50399 PCT/US98/09385 81 Compounds of the subject invention showed a dose response when administered with tetanus toxoid. BFDI (C57B1/6 X DBA/2) female mice (8 per group) were immunized with 0.2 ml of emulsions containing AGP 0.2 pg of tetanus toxoid. A second immunization was administered 21 days post primary immunization. Each mouse was bled 21 days after the second injection. The results are shown in Tables 5 and 6.

T Table Dose response of AGPs in mice immunized with tetanus toxoid.

Material 50 pg 25 pg 10 ipg B27 50 Lpg Vehicle Total Ig T/C Titer Ratio* 3.3 7,000 5.8 12,400 5.3 11,450 3.2 6,800 2150 IgM IgG, IgG2.

T/C

Ratio 13.4 2.1 1.4 4.0 Titer 37,600 6,000 4,000 11,200 2800

T/C

Ratio 4.1 4.5 5.5 1.6 Titer 26,300 28,800 35,100 10,400 6350

T/C

Ratio 150.0 52.0 33.8 12.0 Titer 11,225 3900 2538 900

T/C

Ratio 3.2 7.0 9.9 11.6 Titer 2500 5400 7650 9,000 IgG2b 775 T/C Ratio Experimental Test Titer Vehicle Control Titer.

J 4.

Table 6.

Dose response of AGPs in mice immunized with tetanus toxoid.

Material B12 50 pg B12 25 pg B12 10 pg B12 5 pg B12 1 pg 50 pg 25 pg 10 pg 5 jlg 1 pg IgM Total Ig IgG, IgG2.

IgG2b

T/C*

5.43 3.14 3.71 3.43 1.71 Titer 869 503 594 549 274

T/C

368.55 403.98 248.06 489.92 326.02 Titer 47,543 52,114 32,000 63,200 42,057

T/C

141.22 145.21 81.12 84.11 90.08 Titer 259,429 266,743 149,029 154,514 165,486 165,486 194,971 71,771 189,486 132,343

T/C

16.86 6.81 34.14 73.71 Titer nd 354 143 717 1,548 1,051 219 54 63 160

T/C

499.35 196.92 181.12 352.54 175.81 Titer 12,983 5,120 4,709 9,166 4,571 6,126 4,114 2,194 5,483 2,971 3.14 2.29 2.86 1.71 1.57 503 233.88 30,171 90.08 366 181.91 23,467 106.14 457 170.10 21,943 39.07 274 248.06 32,000 103.15 251 166.56 21,486 72.04 50.05 10.43 2.57 3.00 7.62 235.62 158.23 84.38 210.88 114.27 Vehicle 160 1837 *T/C=Experimental Test Titer Vehicle Control Titer.

nd-not done WO 98/50399 PCT/US98/09385 84 TEST EXAMPLE 2 Stimulation of Antiovalbumin Antibody Production.

BDF 1 female mice (8 per group) were immunized with 0.2 ml of emulsions containing 50 pg of the AGPs 50 ug of ovalbumin. A second immunization was administered 21 days post primary. Each mouse was bled 14 days after the second injection. Antibody titers of immunized mice showing total IgG and IgM as well as titers for the subgroups of IgG including IgG,, IgG 2 a and IgG2b are given in Table 7.

Table 7.

Adjuvant activity in BDF mice immunized with ovalbumin.

Material Total Ig IgM Bll B2 Bl B21 B27

T/C*

0.7 2.5 0.5 1.9 0.5 4.1 0.6 Titer 150 563 119 438 113 925 138 225 Titer 250 175 150 150 250 438 300 188 Vehicle T/C Ratio Experimental Test Titer Vehicle Control Titer WO 98/50399 PCT/US98/09385 Table 7 continued.

Material Bll B2

BI

B21 B27 Vehicle IgG1

T/C*

1.6 5.0 0.5 5.2 0.6 0.6 0.8 Titer 2650 8300 763 8500 1000 950 1275 1650

T/C

1.7 2.5 0.2 0.5 0.1 0.3 0.1 IgG2a Titer 550 825 56 163 25 113 38 325 IgG2b T/C Titer 1.6 375 2.3 550 0.8 188 5.0 1188 0.8 200 16.7 3963 0.5 113 238 T/C Ratio Experimental Test Titer Vehicle Control Titer The AGP compounds of the subject invention when administered to a warmblooded animal with the antigen ovalbumin stimulates the production of antibody to that antigen.

TEST EXAMPLE 3 Generation of a Protective Immune Response to Infectious Influenza.

Mice vaccinated with formalin-inactivated influenza and the AGP compounds of the subject invention mounted a protective immune response to an influenza challenge as well as produced antibody to that antigen. Animals were vaccinated with the antigen and AGP compounds in various carriers. The degree of protection was determined by challenging the mice with intranasal (IN) admininstration of approximately 10 LD 50 infectious influenza A/HK/68. Mortality was assessed for 21 days following the challenge. The number of mice surviving the challenge dose is a direct assessment of the efficacy of the vaccine. For the experiments provided this data does not necessarily correlate with the amount of antibody produced.

WO 98/50399 PCT/US98/09385 86 1) Vaccines were formulated in 0.2% triethanolamine (TEoA)/water solution containing: 1 hemagglutinating unit (HAU) of formalin-inactivated influenza A/HK/68 (FI-Flu), and 50 ig of AGP except the vehicle control vaccines which contained no AGP. ICR mice (10/group) were vaccinated 1 time only. The vaccines were administered by subcutaneous (SQ) injection of 0.1 ml/site at 2 distinct sites near the inguinal lymph nodes for a total of 0.2 ml of vaccine/mouse. Mice (only mice/group) were bled from the orbital plexus 14 days following the vaccination.

Sera was harvested and frozen at -20 0 C until used for enzyme-linked immunosorbent assay (ELISA). All mice were challenged 30 days post vaccination by intranasal (IN) administration of approximately 10 LD 5 s infectious influenza A/HK/68. Mortality was assessed for 21 days following the challenge. Anti-influenza antibody titers obtained from vaccinations with TEoA formulations and corresponding survival rates of mice vaccinated with this formulation are shown in Table 8.

Table 8.

Anti-influenza antibody titers and survival rates of treated mice.

Material Titer-' Percent Total IgG Survival Nonimmune <100 0 Vehicle <100 0 B9 6400 44 1600 B7 200 33 B3 1600 33 B14 6400 44 B15 6400 2) Vaccines were formulated in 2% Squalene solution containing: 1 hemagglutinating unit (HAU) of formalin-inactivated influenza A/HK/68 (FI-Flu), and 25 pig of AGP except the saline and vehicle control vaccines which contained no AGP. BALB/c mice (10/group) were vaccinated 1 time only. The vaccines were administered by subcutaneous (SQ) injection of 0.1 ml/site at 2 distinct sites near the WO 98/50399 PCTIUS98/09385 87 inguinal lymph nodes for a total of 0.2 ml of vaccine/mouse. Mice (only mise/group) were bled from the orbital plexus 14 days following the vaccination.

Sera was harvested and frozen at -20 0 C until used for enzyme-linked immunosorbent assay (ELISA). All mice were challenged 35 days post vaccination by intranasal (IN) administration of approximately 10 LD 5 s infectious influenza A/HK/68. Mortality was assessed for 21 days following the challenge. Anti-influenza antibody titers obtained from vaccinations with the squalene formulations as well as corresponding survival rates of vaccinated animals are shown in Table 9.

Table 9.

Anti-influenza antibody titers and survival rates of treated mice.

Titer' Material Total IgG IgGI IgG 2 IgG2b Percent Survival Nonimmune <100 <100 <100 <100 0 Saline 800 100 800 100 62.5 Vehicle 1600 1600 1600 1600 100 3200 1600 6400 1600 100 1600 3200 3200 400 100 B9 1600 1600 3200 800 87.5 400 400 400 400 62.5 B3 3200 3200 6400 800 87.5 B6 800 800 400 1600 B14 3200 6400 3200 6400 87.5 B28 800 400 400 100 3) The antibody titers and survival rate of vaccinated mice were compared after a primary then a secondary vaccination. Vaccines were formulated in 0.2% TEoA/water solution containing: 1 hemagglutinating unit of formalin-inactivated influenza A/HK/68, 25 pg AGP, except the vehicle control vaccine which contained no AGP. ICR mice (20/group) were administered vaccines by subcutaneous injection of 0.1 ml/site at 2 distinct sites near the inguinal lymph nodes for a total of 0.2 ml of vaccine/mouse. Each group was split into 2 subgroups 35 days after the primary WO 98/50399 PCT/US98/09385 vaccination. One of each subgroup was challenged at this time, the remaining subgroups received a secondary vaccination. Mice (only 5/subgroup) were bled from the orbital plexus 14 days following vaccination (primary or secondary). Sera was harvested and frozen at -20 0 C until used for ELISA. Mice were challenged 35 post primary, or secondary, vaccination by intranasal administration of approximately LDo 0 or 40 LD 5 0 infectious influenza A/HK/68, respectively. Mortality was assessed for 21 days following the challenge. Anti-influenza antibody titers and survival rates of mice post primary and post secondary vaccination are shown in Table Antibody titers as well as survival rates of mice vaccinated a second time were higher.

0 Table Antibody titers and survival rates of treated mice.

Material Nonimmune Vehicle B9 B7 B4 B3 B6 B14 IgG Titer' post 10 post 20 200 100 800 102,400 6400 12,800 1600 25,600 3200 >102,400 800 25,600 3200 102,400 1600 >102,400 1600 102,400 800 51,200 Percent Survival post 10 post 0 0 20 80 60 60 50 70 60 80 33 TEST EXAMPLE 4 The Effect of Fatty Acid Chain Length on Adjuvanticity.

The effect of the length of fatty acid chains RI-R 3 on activity was tested.

Vaccines were formulated in 0.2% TEoA/water solution containing: 1 hemagglutinating unit of formalin-inactivated influenza A/HK/68, and 25 p.g of AGP, except the vehicle control vaccines which contained no AGP. ICR mice WO 98/50399 PCT/US98/09385 89 were vaccinated 1 time only. The vaccines were administered by subcutaneous injection of 0.1 ml/site at 2 distinct sites near the inguinal lymph nodes for a total of 0.2 ml of vaccine/mouse. Mice (only 5 mice/group) were bled from the orbital plexus 14 days following the vaccination. Sera was harvested and frozen at -20°C until used for ELISA. All mice were challenged 35 post vaccination by intranasal administration of approximately 10 LDso infectious influenza A/HK/68. Mortality was assessed for 21 days following the challenge. The length of the fatty acid chain appears to mildly affect biological activity. Results are shown in Tables 11 and 12.

Table 11. 0 Antibody titers and survival rates of treated mice. 0 Titer' Material Chain Length Total IgG IgG, IgG 2 IgG 2 b Percent Survival Nonimmune 200 100 100 800 0 Vehicle 200 100 100 200 11 B18 7 800 800 800 400 1317 8 6400 3200 3200 1600 B316 9 800 1600 100 800 10 3200 200 3200 6400 1314 10 800 1600 100 400 B13 11 1600 800 400 800 B12 12 200 200 100 200 0 1311 14 1600 200 1600 400 00 00 Table 12.

Antibody titers and survival rates of treated mice.o 00 Titer' Material Chain Length Total IgG IgG, IgG 2 IgG 2 b Percent Survival Nonimmune 100 100 50 800 0 Vehicle 100 200 50 100 B8 7 6400 3200 400 1600 B7 9 3200 3200 100 1600 B5 10 800 200 50 400 44 B4 11 3200 400 100 1600 B3 12 1600 1600 50 800 0 BI 14 12,800 6400 1600 15600 WO 98/50399 PCT/US98/09385 92 TEST EXAMPLE The Effect of Variations in the Carbon Chain Length Between the Heteroatom X and the Aglycon Nitrogen Atom on Adjuvanticity.

The length of the carbon chain between X and the aglycon nitrogen atom was extended progressively by a single atom. The effect of lengthening the chain between these two components on adjuvanticity was explored. Vaccines were formulated in 0.2% TEoA/water solution containing: 1 hemagglutinating unit of formalininactivated influenza A/HK/68, and 25 gg of AGP, except the vehicle control vaccines which contained no AGP. ICR mice (10/group) were vaccinated 1 time only. The vaccines were administered by subcutaneous injection of 0.1 ml/site at 2 distinct sites near the inguinal lymph nodes for a total of 0.2 ml of vaccine/mouse.

Mice (only 5 mice/group) were bled from the orbital plexus 14 days following the vaccination. Sera was harvested and frozen at -20 0 C until used for ELISA. All mice were challenged 35 days post vaccination by intranasal administration of approximately 10 LD 50 infectious influenza A/HK/68. Mortality was assessed for 21 days following the challenge. Adjuvant activity appears to lessen as the length of the carbon chain between the heteroatom X and aglycon nitrogen atom increases.

However, depending upon the residues attached to this carbon chain the biologic and metabolic stability of the molecules may be affected. Results are shown in Tables 13.

Table 13.

Antibody titers and survival rates of treated mice.

Titer' Material Carbon Chain Nonimmune Vehicle B19 B21 B22 Total IgG <50 200 12,800 6400 3200 IgG, <50 200 100 800 100 IgG 2 3 <50 50 800 100 3200 IgG 2 b <50 200 6400 1600 200 Percent Survival 0 WO 98/50399 PCT/US98/09385 94 TEST EXAMPLE 6 Cytokine Induction by the AGP Compounds.

The AGP compounds of the subject invention induced cytokines in human whole blood ex vivo culture assays. AGP compounds were solubilized in 10% EtOHwater and diluted to various concentrations. Fifty pl of each dilution were added to 450 pl of whole human blood. Controls were treated with culture media (RPMI).

The reaction mixture was incubated at 37°C for 4 hr with constant mixing on a rotator. Sterile PBS (1.5 ml) was added to the reaction mixture, the cells were centrifuged and the supematents removed for cytokine testing. The concentration of TNF-a and IL- 1 in each supernatent was determined using immunoassay ELISA kits from R&D Systems. Results from these studies are shown in Tables 14-19.

Table 14.

Stimulation of cytokine secretion in an ex vivo assay.

Material Dosage TNF-a IL-1p (pLg) (pg/ml) (pg/ml) B26 20 498.90 33.25 254.94 25.34 75.62 9.89 1 38.85 3.90 B2 20 1338.42 155.07 817.67 114.41 235.32 34.72 1 105.52 14.53 RPMI 2 0 I- WO 98/50399 WO 9850399PCTIUS98/09385 I- Table Stimulation of cytokines in an ex vivo assay.

Material Dosage (ng/mI) B 16 10,000 5000 1000 B13 10,000 5000 1000 B9 10,000 5000 1000 BlO 10,000 5000 1000 B7 10,000 5000 1000 RPMI *THTC-To high to Count TNF-a (pg/mi) 291 277 155 775 716 740 449 247 145 207 127 73 83 57 26 2 IL-ip (pg/mi) 53 39

TH~TC*

187 177 96 84 53 43 61 17 16 14 6 0 WO 98/50399 WO 9850399PCT/US98/09385 96 Table 16.

Stimulation of cytokines in an ex vivo assay.

Material Dosage TNF-at IL-iji (ng/ml) (pg/mi) (pg/mi) B4 10,000 432 213 5000 205 164 1000 94 B3 10,000 567 269 5000 390 342 1000 189 204 10,000 169 79 5000 143 162 1000 43 36 B6 10,000 94 52 5000 59 29 1000 30 13 B14 10,000 249 91 5000 120 71 1000 56 46

RPMI

WO 98/50399 PCT[US98/09385 97 Table 17.

Stimulation of cytokine secretion in an ex vivo assay.

Material Dosage TNF-a IL-1fp (ng/ml) (pg/mi) (pg/mi) BlI 10,000 181 62.3 5000 139 61.7 1000 115 54.5 500 125 55.8 100 127 59.8 B13 10,000 583 282 5000 592 390 1000 478 327 500 411 352 100 302 261 10,000 320 153 5000 280 126 1000 209 94.4 500 183 104 100 133 51.6 B16 10,000 121 41.0 5000 114 34.0 1000 72 19.5 500 55 17.1 B14 10,000 114 24.6 5000 87 19.0 1000 51 10.0 500 49 19.9

RPMI

WO 98/50399 WO 9850399PCT11US98/09385 98 Table 18.

Stimulation of cytokine secretion in an ex vivo assay.

Material Dosage TNF-at IL-iji (ng/ml) (pg/mi) (pg/mi) B2 10,000 100 22.2 5000 75 14.0 1000 38 500 28 8.3 100 6.1 BI 10,000 20 10.0 5000 11 1000 2.8 500 1.1 0 100 0 0 B7 10,000 61 14.7 5000 44 8.3 1000 30 4.3 500 27 3.8 100 10 5.1 B4 10,000 232 66.9 5000 173 66.5 1000 130 32.0 500 116 19.3 100 89 65.2 B3 10,000 433 151.9 5000 316 200.4 1000 229 75.1 500 212 67.9 100 130 35.9 10,000 142 24.1 5000 99 23.0 1000 96 10.5 500 59 16.9 100 33 5.4 RPMI 2 0 WO 98/50399 WO 9850399PCTIUS98/09385 99 Table 19.

Stimulation of cytokine secretion in an ex vivo assay.

Material Dosage TNF-at IL-iIp (ng/ml) (jpg/ml) (pg/mi) B17 10,000 2.8 0 5000 2.2 0 1000 2.6 0.2 B8 10,000 2.8 0 5000 0.7 1000 1.5 0.1 B22 10,000 287 17 5000 11 1.9 1000 2.2 0.1 B28 10,000 198 13 5000 197 13 1000 139 8 B 12 10,000 1017 135 5000 957 153 1000 863 175 RPMI 3.9 0 WO 98/50399 PCT/US98/09385 100 TEST EXAMPLE 7 Stimulation of a Cytotoxic T-lymphocyte Response.

The induction of a cytotoxic T-lymphocyte response after administration of the AGP compounds of the subject invention and a protein antigen was detected by a cytotoxicity assay. Groups of C57BL/6 mice were given a primary immunization subcutaneously (inguinal region) with 25 p.g ovalbumin (OVA) formulated in AGP preparations. The injected volume was 200 il. Twenty-one days later three mice per experimental group were killed and spleens removed and pooled as single cell suspensions and counted.

Spleen cells (75 X 10 6 cells in 3-4 ml media) from the experimental groups were placed in a 25 cm 2 T-flask. Next, 1.0 ml of irradiated (20,000 rads) E.G7 (OVA) cells at 5 X 10 6 /ml were added to the flask. The volume was brought to 10 ml. The cultures were maintained by placing the T-flasks upright in a 37 0 C, 5% CO 2 incubator for four days. On day 4 the surviving cells were recovered from the flasks, washed 1X in fresh media resuspended in 5.0 ml, and counted.

Recovered effector cells were adjusted to 5 X 10 6 viable cells/mi and 100 ul volumes were diluted serially in triplicate in wells of 96 well round-bottom plates (Corning 25850) using 100 l/well of media as a diluent. Next, 100 ul volumes of "Cr-labelled (see below) targets [E.G7 (OVA)-an ovalbumin gene transfected EL-4 cell line] at 1 X 105 cells/ml were added to the wells. Spontaneous release (SR) wells contained 100 pl of targets and 100 pl of media. Maximal release (MR) wells contained 100 pl of targets and 100 ul detergent Tween 20). Effector/target ratios were 50:1, 25:1, 12.5:1. The plates were centrifuged at 400 Xg and incubated at 37°C, 5% CO 2 for 4 hr. After the incubation the well supernatants were collected using a Skatron Supernatant Collection System.

Percent specific lysis= 100 X (Exp.Release

SR)

(MR SR) Target cells, E.G7 (OVA), were labelled with 5 Cr (sodium chromate) as follows. In a total volume of 1.0 ml were mixed 5 X 10 6 target cells and 250 LCi 51 Cr 0 WO 98/50399 PCT/US98/09385 101 in 15 ml conical tube. The cell suspensions was incubated in a 37°C water bath for min., with gentle mixing every 15 min. After incubation the labelled cells were washed 3X by centrifugation and decanting with 15 ml volumes of media. After the third centrifugation the cells were resuspended in 10 ml of fresh media and allowed to stand at room temperature for 30 min. and then centrifuged. The cells were finally resuspended in media at 1 X 10' cells/ml.

Mice immunized according to the procedure above with the AGPs of the subject invention displayed a cytotoxic T-lymphocyte response to the OVA antigen as shown in Table 20. Table 0 Cytotoxic T-lymphocyte response of treated cells.

Cytotoxicity Material Bll B12 B13 B16 B17 B18 B14 B28 B27 50:1 14 13 28 58 42 39 36

E:T

25:1 8 7 15 49 29 26 20 12.5:1 4 45 36 1 WO 98/50399 PCT/US98/09385 102 Table 20 continued.

Cytotoxicity Material B1 B3 B4 B7 B8 B9 B6 B2 B26 B22 B21 B19 Vehicle Control 50:1 34 65 72 28 57 36 65 65 54 21 65 14 58 38 59 36

E:T

25:1 24 54 66 18 44 20 56 55 41 12 55 8 42 26 42 25 12.5:1 42 11 29 38 36 37 6 43 4 31 33 13 TEST EXAMPLE 8 Generation of Serum and Mucosal Antibody Titers to Tetanus-toxoid.

The AGPs of the subject invention elicited both a serum and mucosal immune response to purified tetanus toxoid when administered intranasally. Groups of BALB/c mice were given a primary immunization (1 o) intranasally with 10 Pg tetanus toxoid (TT) 20 ig AGP formulated in an aqueous formulation (AF) in a WO 98/50399 PCT[US98/09385 103 volume of 20 pl. A secondary immunization was given 14 days later and a tertiary immunization (3o) identical in composition to the first and second was administered 14 days later. Mice were bled on day 21 (day 7 post and day 38 (day post 30) and day 48 (day 20 post Vaginal wash/fecal extract samples were taken on day 7 post 20 and day 7 post 3 Serum and wash samples were assayed for anti-TT antibody by standard ELISA methods. Results of these assays are shown in Tables 21 and 22 below.

The aqueous formulation comprises the AGPs of the subject invention and one or more surfactants. Surfactants useful in an aqueous composition include glycodeoxycholate, deoxycholate, sphingomyelin, sphingosine, phosphatidylcholine, 1,2-Dimyristoyl-sn-glycero-3-phosphoethanolamine, L-a-phosphatidylethanolamine, and 1, 2 -Dipalmitoyl-sn-glycero-3-phosphocholine, or a mixture thereof. The aqueous formulation used in this example comprises the surfactant 1,2 dipalmitoyl-sn-glycero- 3-phosphocholine (DPPC) and was prepared as follows: briefly; a 4 mg/ml solution of DPPC was prepared in ethanol. An aliquot of the ethanol solution is added to the dried AGPs and swirled gently to wet the AGP. The ethanol is removed by blowing a stream of filtered nitrogen gently over the vial. Water for Injection is added and the suspension is sonicated 10 min. at 60°C until clear. The resulting aqueous formulation contains approximately 118 ig/ml DPPC, has particles of around and was filter sterilized.

WO 98/50399 PCT/US98/09385 104 Table 21.

Anti-tetanus toxoid antibody titers in treated mice.

Vaginal IgG Anti-Tetanus Toxoid Titer-' Wash Fecal Extract IgA IgG IgA Vaginal IgG Material B19 B4 B3 B22

PBS

Normal Sera 2° 30 20 30 20 800 6400 6400 6400 50 400 800 6400 6400 50 200 400 1600 3200 25 1600 400 1600 6400 25 3200 800 3200 3200 50 1600 1600 6400 6400 50 400 800 800 3200 25 <25 <25 <25 <25 <25 <25 <25 <25 <25 <25 3° 20 200 3200 6400 100 6400 12,800 25 3200 6400 50 3200 12,800 100 3200 6400 100 3200 6400 50 1600 6400 <25 <25 <25 <25 Table 22.

Serum anti-tetanus toxoid antibody titers in treated animals.

00 Anti-Tetanus Toxoid Titer 1 Serum Pools Material IgG, IgG 2 a IgA d21 d38 d48 d21 d38 d48 d21 d38 d48 IM* 8M 8M 512K 4M 4M 12,800 102,400 102,400 2M 8M 8M 512K 1M 2M 12,800 51,200 25,600 B19 2M 4M 4M 64K# 256K 128K 6,400 25,600 12,800 B4 IM 8M 8M IM 2M 2M 25,600 102,400 102,400

U

2M 8M 8M 512K 2M 2M 25,600 102,400 102,400 B3 512K 4M 8M 512K 2M 2M 12,800 51,200 51,200 B22 1M 2M 4M 64K 256K 256K 6,400 25,600 25,600 PBS 1,000 16K 16K 1,000 1,000 1,000 200 200 200 Normal 200 200 200 100 100 100 200 200 200 Sera 6 #K=10 3 00 01 WO 98/50399 PCT/US98/09385 106 Intranasal administration of TT formulated in AGP-AF induced both an antigen specific humoral immune response (Table 22 and a mucosal immune response (Table 21) to that antigen.

TEST EXAMPLE 9 Stimulation of an Immune Response to Hepatitis B Surface Antigen by Intranasal Administration Mice administered hepatitis B surface antigen (HBsAg) intranasally with the compounds of the subject invention produced serum IgG and IgA titers to that antigen. Secretory IgA was detected in vaginal washes and the induction of a cytotoxic T-lymphocyte response was detected by a cytotoxicity assay.

Groups of BALB/c mice were given a primary immunization (1 o) intranasally with 2.5 ltg HBsAg 10 ug AGP-AF in a volume of 20 ll. AGP-AF was prepared as in TEST EXAMPLE 8. Twenty-one days later mice were given a secondary immunization (20) of 7.5 ig HBSAG 10 tg AGP-AF intranasally in 20 pl volume.

A tertiary immunization (3 identical in composition to the secondary immunization was administered 28 days after the secondary immunization. Assays were conducted to detect cytotoxic T-lymphocyte activity at 16 days post secondary immunization (dl6 post and 8 days post tertiary immunization (d8 post 3 Serum and mucosal antibody titers were assessed at 22 days post secondary immunization (d22 post and 21 days post tertiary immunization (d21 post 30). Antibody assays were conducted by standard ELISA methods. Cytotoxicity assays were conducted as described in TEST EXAMPLE 7. Results from this experiment are shown in Tables 23-26.

WO 98/50399 WO 9850399PCTIUS98/09385 107 Table 23.

Cytotoxic T-lymnphocyte response of treated cells.

Cytotoxicity (d16, post

E/T

Material 50:1 25:1 12.5:1 6.25:1 B 19 B4 B3 B22 Vehicle Normal Cells Table 24.

Cytotoxic T-lymphocyte response of treated cells.

Cytotoxicity (d8, post

E/T

Material 50:1 25:1 12.5:1 6.25:1 B19 B4 B3 B22 Vehicle Normal Cells 30 19 56 42 71 54 23 54 44 22 13 5 2 7 WO 98/50399 PCT/US98/09385 108 Table Anti-hepatitis antibody titers in treated mice.

Anti HBsAg Titer'* Material IgG, IgG2. IgA B25 256K# 500K 3,200 256K 500K 6,400 B19 500K 64K 1,600 B4 500K 1000K 6,400 B3 1000K 500K 6,400 B5 256K 500K 3,200 B22 256K 64K 1,600 Vehicle <2K <2K <200 day 22 post 20, #K=10 3 Table 26.

Anti-hepatitis antibody titers in treated mice.

Anti HBsAg Titer-' Material IgG, IgG 2 a IgA 1000K# 1000K 25,600 B15 2000K 2000K 25,600 B19 2000K 500K 12,800 B4 1000K 2000K 25,600 B3 1000K 1000K 25,600 500K 1000K 12,800 B22 500K 500K 12,800 Vehicle <2K <2K <200 day 21 post 30, #K=10 3 Groups of BALB/c mice were immunized with 2.5 ig HBsAg 10 [g AGP- AF intranasally and boosted intranasally with 7.5 pg HBsAg 10 utg AGP-AF 21 days later. Vaginal samples were collected 10 days after the booster immunization and assayed for anti-HBsAg antibody. Results of this assay are shown in Table 27.

11 WO 98/50399 PCT/US98/09385 109 Table 27.

Vaginal Wash Anti-HBsAg Titer' Material IgG IgA B25 100 800 50 3200 B19 <50 400 B4 1600 6400 B3 800 1600 B5 1600 1600 B22 100 800 Vehicle <50 The intranasal administration of HBsAg with the compounds of the subject invention stimulated both a humoral and cellular immune response to that antigen.

Intranasal immunization with the antigen formulated in AGP-AF induced a cytotoxic T-lymphocyte response (Table 23-24) and antigen specific humoral (Table 25 and 26) and mucosal (Table 27) immune responses.

TEST EXAMPLE Generation of a Protective Immune Response to Influenza Mice immunized intranasally with FLUSHIELD influenza vaccine containing hemagglutinin antigen and the AGPs of the subject invention produced both IgG and IgA which were recovered in vaginal washes. Immunized mice were also protected from subsequent influenza challenge.

ICR mice were immunized three times at 21 day intervals intranasally with FLUSHIELD influenza vaccine (Wyeth-Lederle) containing 0.3 tg hemagglutinin antigen (HA) 10 tg AGP-AF or recombinant E. coli heat labile enterotoxin (LT).

AGP-AF was prepared as in TEST EXAMPLE 8. LT was solubilized in saline at 1 g/ml. Vaginal washes were collected 14 days after the second and third WO 98/50399 PCT/US98/09385 110 immunization. Serum samples were collected 14 days after the third immunization.

Mice were challenged with 10 LDso (lethal dose 50) of infectious influenza A/HK/68 thirty-five days after the final immunization and monitored for mortality. Tables 28 and 29 show the results of assays conducted by standard ELISA methods to detect anti-influenza antibody titers in vaginal washes and sera.

Table 28.

Vaginal Wash Samples Material Nonimmune Vehicle B19 B3 B22

LT

IgA Secondary <20 80 1280 320 1280 640 2560 Tertiary <20 160 1280 5120 2560 2560 2560 IgG Secondary <20 160 640 1280 1280 320 2560 Tertiary <20 160 2560 1280 1280 640 640 Percent Protection 22 100 100 100 Table 29.

Material Nonimmune Vehicle B19 B3 B22

LT

Total IgG <400 102,400 2819,200 819,200 2819,200 819,200 2819,200 Serum Titers IgG, IgG 2 <400 <400 256,000 12,800 102,400 819,200 51,200 102,400 51,200 819,200 51,200 102,400 2819,20 2819,200 0 IgG2b <400 102,400 2819,200 819,200 2819,200 819,200 2819,200 Percent Protection 22 100 100 100 WO 98/50399 PCT/US98/09385 111 These data demonstrate that AGPs in AF when administered intranasally act as a mucosal adjuvants causing the production of IgA at mucosal sites. Increased protection is also induced against an upper respiratory pathogen which invades through the mucosa.

TEST EXAMPLE 11 Generation of Immune Responses from Stable Emulsion Formulations.

The AGP compounds of the subject invention stimulated both humoral and cytotoxic T-lymphocyte responses when formulated in a stable emulsion AGPs were tested at 25 tg dose levels to adjuvantize Hepatitis B surface antigen (HBsAg) for the induction of CTL and antibody responses. BALB/c mice were immunized subcutaneously with 2.0 gg HBsAg plus 25 pig of AGP/SE on day 0 and day 21. The CTL assay was conducted as in TEST EXAMPLE 7. The AGPs were formulated in a stable emulsion (SE) and the compositions were designated AGP-SE. Methods for preparing the stable emulsion containing 10% v/v squalene, 0.091% w/v PLURONIC- F68 block copolymer, 1.909% w/v egg phosphatidyl choline, 1.8% v/v glycerol, 0.05% w/v a tocopherol, 10% ammonium phosphate buffer and 78.2% v/v Water for Injection should be readily apparent to one skilled in the art. The emulsion was homogenized to a particle size of <0.2 gm. Table 30 shows the AGPs of the subject invention induced a cytotoxic T-lymphocyte response to HBsAg.

WO 98/50399 PCT/US98/09385 112 Table Cytotoxic T-lymphocyte response of treated cells.

Cytotoxicity

E:T

Material Day 50:1 25:1 12.5:1 6.25:1 B25 dl7,post 1 27 12 9 B19 74 48 34 24 B3 28 15 9 B22 42 24 17 7 Vehicle-SE 32 16 9 6 d16, post 2° 49 28 20 13 B19 73 62 42 31 B3 81 47 32 22 B22 78 69 58 39 Vehicle-SE 38 23 14 8 The results of the antibody titer to HBsAg are shown on Table 31. Sera from bleeds taken on day 28 post 20 were titered on ELISA plates coated with either HBsAg or a 28 amino acid peptide (p72) which contains B-cell epitopes found in the S-antigen region, residues 110-137, of the HBsAg.

Table 31.

Anti-HBsAg titer of treated mice.

Anti-HBsAg Titer' HBsAg p72-Peptide Material IgG, IgG 2 IgG, IgG,, 2048 K* 2048 K 128 K 64 K B19 1024 K 1024 K 64 K 128 K B3 512 K 1024 K 16K 128 K B22 1024 K 1024 K 128 K 128 K Vehicle-SE 1024 K 64 K 64 K 4K WO 98/50399 PCTIUS98/09385 113 AGP-SE treated mice displayed both humoral (Table 31) and cytotoxic Tlymphocyte (Table 30) responses to the hepatitis B surface antigen. Of interest, AGP- SE treated mice in serum displayed a vigorous IgG,, specific antibody titer detected by both antigens, whereas the vehicle-SE induced only a modest IgG2a response.

It is understood that the foregoing examples are merely illustrative of the present invention. Certain modifications of the compositions and/or methods employed may be made and still achieve the objectives of the invention. Such modifications are contemplated as within the scope of the claimed invention.

WO 98/50399 PCT/US98/09385 114 References Bulusu, Waldstatten, Hildebrandt, Schtitze, E. and G. Schulz (1992) Cyclic Analogues of Lipid A: Synthesis and Biological Activities, J. Med.

Chem. 35: 3463-3469.

Ikeda, Asahara, T. and K. Achiwa (1993) Synthesis of Biologically Active Nacylated L-serine-Containing Glucosamine-4-Phosphate Derivatives of Lipid A, Chem. Pharm. Bull. 41(10): 1879-1881.

Miyajima, Ikeda, K. and K. Achiwa (1996) Lipid A and Related Compounds XXXI. Synthesis of Biologically Active N-Acylated L-Serine-Containing D- Glucosamine 4-Phosphate Derivatives of Lipid A, Chem. Pharm. Bull. 44(12): 2268-2273.

Shimizu, Akiyama, Masuzawa, Yanagihara, Nakamoto, Takahashi, Ikeda, K. and K. Achiwa (1985) Antitumor Activity and Biological Effects of Chemically Synthesized Monosaccharide Analogues of Lipid A in Mice.

Chem. Pharm. Bull. 33(10): 4621-4624.

Shimizu, Sugiyama, Iwamoto, Yanagihara, Asahara, Ikeda, K. and K. Achiwa (1994) Biological Activities of Chemically Synthesized N-acylated Serine-linked Lipid A Analog in Mice, Int. J. Immunopharmac., 16(8): 659- 665.

Shimizu, lida, Iwamoto, Yanagihara, Ryoyama, Asahara, Ikeda, K. and K. Achiwa (1995) Biological Activities and Antitumor Effects of Synthetic Lipid A Analogs Linked N-Acylated Serine, Int. J.

Immunopharmac., 17(5): 425-431.

Claims (26)

1. 2. The compound of claim 1, wherein R is carboxy. WO 98/50399 PCTIUS98/09385 116 1
2. 3. The compound of claim 2, wherein X is O; Y is O; n, m, p and q are 0; R,, 2 R 2 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R, and R 7 are H; 3 R 8 is phosphono; R 9 is H; R 2 and R 3 are each attached to a stereogenic center 4 having an R configuration; and R, is attached to a stereogenic center having an S configuration. 1
3. 4. The compound of claim 2, wherein X is O; Y is O; n, m, p and q are 0; R,, 2 R 2 and R 3 are normal fatty acyl residues having 12 carbon atoms; R 4 R 5 and R 7 are H; 3 R 8 is phosphono; R 9 is H; R, and R 3 are each attached to a stereogenic center 4 having an R configuration; and R 5 is attached to a stereogenic center having an S configuration. 1
4. 5. The compound of claim 2, wherein X is O; Y is O; n, m, p and q are 0; R,, 2 R 2 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R 5 and R 7 are H; 3 R, is phosphono; R 9 is H; R 2 and R 3 are each attached to a stereogenic center 4 having an R configuration; and R 5 is attached to a stereogenic center having an R configuration. 1 6. The compound of claim 2, wherein X is O; Y is O; n, m, p and q are 0; R,, 2 R 2 and R 3 are normal fatty acyl residues having 8 carbon atoms; R 4 R 5 and R 7 are H; 3 R 8 is phosphono; R 9 is H; Ri, R, and R 3 are each attached to a stereogenic center 4 having an R configuration; and R 5 is attached to a stereogenic center having an S configuration. 1 7. The compound of claim 1, wherein R 6 is H. 1 8. The compound of claim 7, wherein X is O; Y is O; n is 2; m, p and q are 0; 2 R, and R 3 are normal fatty acyl residues having 14 carbon atoms; R 4 R 5 and R 7 are 3 H; R 8 is phosphono; R, is H; and R 2 and R 3 are each attached to a stereogenic 4 center having an R configuration. WO 98/50399 PCT/US98/09385 117 1 9. The compound of claim 7, wherein X is O; Y is O; n is 1, m and p are 0; q 2 is R, and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 and R, 3 are H; R 7 is carboxy; R 8 is phosphono; R, is H; and R, and R3 are each attached to 4 a stereogenic center having an R configuration. 1
5. 10. The compound of claim 7, wherein X is O; Y is O; m, n, p and q are 0; R,, 2 R, and R3 are normal fatty acyl residues having 14 carbon atoms; R 4 R, and R 7 are H; 3 R 8 is phosphono; R 9 is H; and R, and R 3 are each attached to a stereogenic center 4 having an R configuration. 1
6. 11. The compound of claim 7, wherein X is O; Y is O; m, n, p and q are 0; R,, 2 R, and R 3 are normal fatty acyl residues having 10 carbon atoms; R4, R, and R, are H; 3 R 8 is phosphono; R9 is H; and R, and R 3 are each attached to a stereogenic center 4 having an R configuration. 1 12. The compound of claim 7, wherein X is O; Y is O; m, p and q are 0; n is 2 1; R 2 and R3 are normal fatty acyl residues having 14 carbons; R 4 R5 and R 7 are H; 3 R, is phosphono; R 9 is H; and R, and R3 are each attached to a stereogenic center 4 having an R configuration. 1 13. The compound of claim 1, wherein R 6 is hydroxy. 1 14. The compound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is 2 1; R, and R 3 are normal fatty acyl residues having 12 carbon atoms; R, and R, are 3 H; R 7 is H; R, is phosphono; and R9 is H; RI, R2 and R 3 are each attached to a 4 stereogenic center having an R configuration; and R, is attached to a stereogenic center having an S configuration. WO 98/50399 PCT/US98/09385 118 1 15. The compound of claim 13, wherein X is O; Y is O; m and q are 0; n and 2 p are 1; R 2 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R 3 and R 7 are H; R 8 is phosphono; R, is H; R 2 and R 3 are each attached to a 4 stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an S configuration. 1 16. The compound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is 2 2; R2 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R 5 and R 7 3 are H; R, is phosphono; R 9 is H; R 2 and R 3 are each attached to a stereogenic 4 center having an R configuration; and R 5 is attached to a stereogenic center having an S configuration. 1 17. The compound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is 2 1; R, R2 and R 3 are normal fatty acyl residues having 14 carbon atoms; R4, R5 and R7 3 are H; Rg is phosphono; R9 is H; RI, R2 and R3 are each attached to a stereogenic 4 center having an R configuration; and R 5 is attached to a stereogenic center having an R configuration. 1 18. The compound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is 2 1; R 2 and R 3 are normal fatty acyl residues having 14 carbon atoms; R4, R5 and R 7 3 are H; R8 is phosphono; R 9 is H; R2 and R 3 are each attached to a stereogenic 4 center having an R configuration; and R 5 is attached to a stereogenic center having an S configuration. 1 19. The compound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is 2 1; RI, R2 and R3 are normal fatty acyl residues having 11 carbon atoms; R 4 R5 and R, 3 are H; R, is phosphono; R9 is H; R2 and R 3 are each attached to a stereogenic 4 center having an R configuration; and R5 is attached to a stereogenic center having an S configuration. WO 98/50399 PCT/US98/09385 119 1 20. The compound of claim 13, wherein X is O; Y is O; m, n and q are 0; p is 2 1; R 2 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R 5 and R 7 3 are H; R, is phosphono; R 9 is H; R, and R3 are each attached to a stereogenic 4 center having an R configuration; and R, is attached to a stereogenic center having an S configuration. 1 21. The compound of claim 1, wherein X is O; Y is O; m, n, p and q are 0; R,, 2 R, and R 3 are normal fatty acyl residues having 10 carbon atoms; R4 and R 5 are H; R6 3 is amino carbonyl; R 7 is H; R 8 is phosphono; and R 9 is H; R, and R 3 are each 4 attached to a stereogenic center having an R configuration; and Rs is attached to a stereogenic center having an S configuration. WO 98/50399 PCT/US98/09385 2 3 .0 120
7. 22. A method for enhancing the immune response of a mammal comprising administering to the mammal an effective amount of a compound having the following structure: .1. (C4) (C, 4 (C 14 wherein, X is selected from the group consisting of O and S; Y is selected from the group consisting of O and NH; n, m, p and q are integers from 0 to 6; R 2 and R 3 are normal fatty acyl residues having from about 7 to 16 carbon atoms; R4 and R, are the same or different and are selected from the group consisting of H and methyl; R and R 7 are the same or different and are selected from the group consisting of H, hydroxy, alkoxy, phosphono, phosphonooxy, sulfo, sulfooxy, amino, mercapto, cyano, nitro, formyl and carboxy, and esters and amides thereof; R, and R, are the same or different and are selected from the group consisting of phosphono and H, and at least one of R, and R, is phosphono.
8. 23. The method of claim 22, wherein R of said compound is carboxy. WO 98/50399 PCT/US98/09385 121 1 24. The method of claim 23, wherein said compound has the following 2 structure: X is O; Y is O; n, m, p and q are 0; RI, R, and R 3 are normal fatty acyl 3 residues having 10 carbon atoms; R 4 and R 5 are H; R7 is H; R 8 is phosphono; R 9 is H; 4 R, and R 3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an S configuration. 1 25. The method of claim 23, wherein said compound has the following 2 structure: X is O; Y is O; n, m, p and q are 0; R 2 and R 3 are normal fatty acyl 3 residues having 12 carbon atoms; R 4 and R 5 are H; R 7 is H; R 8 is phosphono; R 9 is H; 4 R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R, is attached to a stereogenic center having an S configuration. 1 26. The method of claim 23, wherein said compound has the following 2 structure: X is O; Y is O; n, m, p and q are 0; R 2 and R 3 are normal fatty acyl 3 residues having 10 carbon atoms; R 4 R 5 and R 7 are H; R 8 is phosphono; R 9 is H; R,, 4 R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R is attached to a stereogenic center having an R configuration. 1 27. The method of claim 23, wherein said compound has the following 2 structure: X is O; Y is O; n, m, p and q are 0; R 2 and R 3 are normal fatty acyl 3 residues having 8 carbon atoms; R 4 R, and R 7 are H; R, is phosphono; R, is H; R 2 4 and R 3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an S configuration. 1 28. The method of claim 22, wherein R6 of said compound is H. 1 29. The method of claim 28, wherein said compound has the following 2 structure: X is O; Y is O; n is 2; m, p and q are 0; R, and R3 are normal fatty acyl 3 residues having 14 carbon atoms; R 4 R5 and R, are H; R 8 is phosphono; R9 is H; and 4 R2 and R3 are each attached to a stereogenic center having an R configuration. WO 98/50399 PCT/US98/09385 122 1 30. The method of claim 28, wherein said compound has the following 2 structure: X is O; Y is O; n is 1, m and p are 0; q is 1; R 2 and R 3 are normal fatty 3 acyl residues having 10 carbon atoms; R 4 and R, are H; R 7 is carboxy; R 8 is 4 phosphono; R 9 is H; and Ri, R, and R 3 are each attached to a stereogenic center having an R configuration. 1 31. The method of claim 28, wherein said compound has the following 2 structure: X is O; Y is O; m, n, p and q are 0; R, and R 3 are normal fatty acyl 3 residues having 14 carbon atoms; R4, R5 and R7 are H; R 8 is phosphono; R 9 is H; and 4 R, and R3 are each attached to a stereogenic center having an R configuration. 1 32. The method of claim 28, wherein said compound has the following 2 structure: X is O; Y is O; m, n, p and q are 0; R2 and R 3 are normal fatty acyi 3 residues having 10 carbon atoms; R4, R 5 and R 7 are H; R 8 is phosphono; R 9 is H; and 4 Ri, R2 and R3 are each attached to a stereogenic center having an R configuration. 1 33. The method of claim 28, wherein said compound has the following 2 structure: X is O; Y is O; m, p and q are 0; n is 1; RI, R2 and R3 are normal fatty acyl 3 residues having 14 carbons; R4, R, and R, are H; R8 is phosphono; R 9 is H; and R2 4 and R 3 are each attached to a stereogenic center having an R configuration. 1 34. The method of claim 22, wherein R6 of said compound is hydroxy. 1 35. The method of claim 34, wherein said compound has the following 2 structure: X is O; Y is O; m, n and q are 0; p is 1; R2 and R 3 are normal fatty acyl 3 residues having 12 carbon atoms; R4 and R, are H; R7 is H; R, is phosphono; and R, is 4 H; Ri, R 2 and R3 are each attached to a stereogenic center having an R configuration; and R5 is attached to a stereogenic center having an S configuration. WO 98/50399 PCT/US98/09385 123 1 36. The method of claim 34, wherein said compound has the following 2 structure: X is O; Y is O; m and q are 0; n and p are 1; R2 and R 3 are normal fatty 3 acyl residues having 10 carbon atoms; R 4 R 5 and R 7 are H; R 8 is phosphono; R 9 is H; 4 R 1 R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an S configuration. 1 37. The method of claim 34, wherein said compound has the following 2 structure: X is O; Y is O; m, n and q are 0; p is 2; R 2 and R3 are normal fatty acyl 3 residues having 10 carbon atoms; R 4 R5 and R7 are H; R, is phosphono; R 9 is H; R,, 4 R, and R3 are each attached to a stereogenic center having an R configuration; and R is attached to a stereogenic center having an S configuration. 1 38. The method of claim 34, wherein said compound has the following 2 structure: X is O; Y is O; m, n and q are 0; p is 1; R 2 and R 3 are normal fatty acyl 3 residues having 14 carbon atoms; R 4 Rs and R7 are H; R, is phosphono; R, is H; R,, 4 R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R, is attached to a stereogenic center having an R configuration. 1 39. The method of claim 34, wherein said compound has the following 2 structure: X is O; Y is O; m, n and q are 0; p is 1; R 2 and R 3 are normal fatty acyl 3 residues having 14 carbon atoms; R 4 R, and R7 are H; R 8 is phosphono; R 9 is H; R,, 4 R2 and R3 are each attached to a stereogenic center having an R configuration; and is attached to a stereogenic center having an S configuration. 1 40. The method of claim 34, wherein said compound has the following 2 structure: X is O; Y is O; m, n and q are 0; p is 1; RI, R 2 and R 3 are normal fatty acyl 3 residues having 11 carbon atoms; R4, Rs and R, are H; R 8 is phosphono; R, is H; R,, 4 R2 and R3 are each attached to a stereogenic center having an R configuration; and R is attached to a stereogenic center having an S configuration. WO 98/50399 PCT/US98/09385 124 1 41. The method of claim 34, wherein said compound has the following 2 structure: X is O; Y is O; m, n and q are 0; p is 1; R 2 and R 3 are normal fatty acyl 3 residues having 10 carbon atoms; R 4 R 5 and R 7 are H; R 8 is phosphono; R, is H; R,, 4 R, and R 3 are each attached to a stereogenic center having an R configuration; and R is attached to a stereogenic center having an S configuration. 1 42. The method of claim 22, wherein said compound has the following 2 structure: X is O; Y is O; m, n, p and q are 0; R 2 and R 3 are normal fatty acyl 3 residues having 10 carbon atoms R 4 and R 5 are H; R6 is amino carbonyl; R 7 is H; R 8 4 is phosphono; R 9 is H; R 2 and R3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an S 6 configuration. WO 98/50399 PCT/US98/09385 125
9. 43. A vaccine composition comprising a compound having the following structure: r (C, 4 (C, 4 (C 4 wherein, X is selected from the group consisting of O and S; Y is selected from the group consisting of O and NH; n, m, p and q are integers from 0 to 6; RI, R 2 and R 3 are the same or different and are normal fatty acyl residues having from about 7 to 16 carbon atoms; R, and R, are the same or different and are selected from the group consisting of H and methyl; R. and R, are the same or different and are selected from the group consisting of H, hydroxy, alkoxy, phosphono, phosphonooxy, sulfo, sulfooxy, amino, mercapto, cyano, nitro, formyl and carboxy, and esters and amides thereof; R, and R are the same or different and are selected from the group consisting of phosphono and H, and at least one of Ra and R, is phosphono, an antigen and a suitable carrier.
10. 44. The composition of claim 43, wherein said composition comprises said compound where PR is carboxy. WO 98/50399 PCT/US98/09385 126 1 45. The composition of claim 44, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n, m, p and q are 0; R 2 3 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R 5 and R 7 are H; R 8 4 is phosphono; R 9 is H; R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an S 6 configuration. 1 46. The composition of claim 44, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n, m, p and q are 0; R, 3 and R 3 are normal fatty acyl residues having 12 carbon atoms; R 4 R, and R 7 are H; R 8 4 is phosphono; R 9 is H; R, and R 3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an S 6 configuration. 1 47. The composition of claim 44, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n, m, p and q are 0; R, 3 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R 5 and R 7 are H; R 8 4 is phosphono; R 9 is H; RI, R, and R 3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an R 6 configuration. 1 48. The composition of claim 44, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n, m, p and q are 0; R 2 3 and R 3 are normal fatty acyl residues having 8 carbon atoms; R 4 R 5 and R 7 are H; R 8 4 is phosphono; R 9 is H; Ri, R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an S 6 configuration. 1 49. The composition of claim 43, wherein said composition comprises said 2 compound where R 6 is H. WO 98/50399 PCT/US98/09385 127 1
11. 50. The composition of claim 49, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n is 2; m, p and q are 0; R,, 3 R 2 and R 3 are normal fatty acyl residues having 14 carbon atoms; R 4 R 5 and R, are H; 4 R, is phosphono; R 9 is H; and R 2 and R3 are each attached to a stereogenic center having an R configuration. 1
12. 51. The composition of claim 49, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n is 1, m and p are 0; q is 3 1; R, and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 and R5 are 4 H; R 7 is carboxy; R, is phosphono; R9 is H; and R, and R 3 are each attached to a stereogenic center having an R configuration. 1 52. The composition of claim 49, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n, p and q are 0; R, 3 and R3 are normal fatty acyl residues having 14 carbon atoms; R 4 R 5 and R 7 are H; R 8 4 is phosphono; R 9 is H; and R 2 and R 3 are each attached to a stereogenic center having an R configuration. 1 53. The composition of claim 49, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n, p and q are 0; Ri, R, 3 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R5 and R7 are H; R 8 4 is phosphono; R 9 is H; and R2 and R 3 are each attached to a stereogenic center having an R configuration. 1
13. 54. The composition of claim 49, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, p and q are 0; n is 1; R,, 3 R 2 and R3 are normal fatty acyl residues having 14 carbons; R 4 R 5 and R 7 are H; R, is 4 phosphono; R 9 is H; and R, and R3 are each attached to a stereogenic center having an R configuration. 1
14. 55. The composition of claim 43, wherein composition comprises said 2 compound where R 6 is hydroxy. WO 98/50399 PCT/US98/09385 128 1
15. 56. The composition of claim 55, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n and q are 0; p is 1; R,, 3 R, and R 3 are normal fatty acyl residues having 12 carbon atoms; R 4 and R 5 are H; R 7 4 is H; R, is phosphono; and R 9 is H; R, and R 3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an 6 S configuration. 1 57. The composition of claim 55, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m and q are 0; n and p are 3 1; R 2 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R 5 and R 7 4 are H; R 8 is phosphono; R 9 is H; R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an 6 S configuration. 1 58. The composition of claim 55, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n and q are 0; p is 2; R,, 3 R, and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R 5 and R 7 are H; 4 R 8 is phosphono; R 9 is H; R 2 and R 3 are each attached to a stereogenic center having an R configuration; and Rs is attached to a stereogenic center having an S 6 configuration. 1
16. 59. The composition of claim 55, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n and q are 0; p is 1; R,, 3 R 2 and R 3 are normal fatty acyl residues having 14 carbon atoms; R 4 R 5 and R 7 are H; 4 R 8 is phosphono; R 9 is H; R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R, is attached to a stereogenic center having an R 6 configuration. WO 98/50399 PCT/US98/09385 129 1 60. The composition of claim 55, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n and q are 0; p is 1; R,, 3 R and R 3 are normal fatty acyl residues having 14 carbon atoms; R, R, and R, are H; 4 R, is phosphono; R, is H; R, and R 3 are each attached to a stereogenic center having an R configuration; and R, is attached to a stereogenic center having an S 6 configuration. 1 61. The composition of claim 55, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n and q are 0; p is 1; R,, 3 R 2 and R 3 are normal fatty acyl residues having 11 carbon atoms; R 4 Rs and R7 are H; 4 R, is phosphono; R% is H; R, and R 3 are each attached to a stereogenic center 5 having an R configuration; and R, is attached to a stereogenic center having an S S. 6 configuration. 1 62. The composition of claim 55, wherein said composition comprises said 2 compound having the following structure: X is 0; Y is m, n and q are 0; p is 1; R,, 3 R 2 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R, and R 7 are H; 4 R, is phosphono; R, is H; R 2 and R 3 are each attached to a stereogenic center 5 having an R configuration; and R, is attached to a stereogenic center having an S 6 configuration. 1 63. The composition of claim 55, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n, p and q are 0; R, 3 and R 3 are normal fatty acyl residues having 10 carbon atoms; RP and R 5 are H; RP is 4 amino carbonyl; R, is H; R, is phosphono; R, is H; R, and R 3 are each attached to a stereogenic center having an R configuration; and R, is attached to a stereogenic 6 center having an S configuration. WO 98/50399 PCT/US98/09385 130
17. 64. A pharmaceutical composition comprising a compound having the following structure: OR, 0 m N q R 7 SNH R4 (CHOp R 1 0 R6 OR3 R 2 0 (C 1 4) ((C1 4 (C 1 4) wherein, X is selected from the group consisting of 0 and S; Y is selected from the group consisting of 0 and NH; n, m, p and q are integers from 0 to 6; R 2 and R, are normal fatty acyl residues having from about 7 to 16 carbon atoms; R 4 and R 5 are the same or different and are selected from the group consisting of H and methyl; R 6 and R, are the same or different and are selected from the group consisting of H, hydroxy, alkoxy, phosphono, phosphonooxy, sulfo, sulfooxy, amino, mercapto, cyano, nitro, formyl and carboxy, and esters and amides thereof; R 8 and R 9 are the same or different and are selected from the group consisting of phosphono and H, and at least one of Rs and R, is phosphono, and a pharmaceutically acceptable carrier. The composition of claim 64, wherein said composition comprises said compound where R 6 is carboxy. WO 98/50399 PCT/US98/09385 131 1 66. The composition of claim 65, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n, m, p and q are 0; RI, R, 3 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R, and R 7 are H; R, 4 is phosphono; R 9 is H; RI, R, and R3 are each attached to a stereogenic center having an R configuration; and R5 is attached to a stereogenic center having an S 6 configuration. 1 67. The composition of claim 65, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n, m, p and q are 0; Ri, R, 3 and R 3 are normal fatty acyl residues having 12 carbon atoms; R 4 Rs and R 7 are H; R 8 4 is phosphono; R 9 is H; R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R5 is attached to a stereogenic center having an S 6 configuration. 1 68. The composition of claim 65, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n, m, p and q are 0; R, 3 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 R, and R7 are H; R 8 4 is phosphono; R, is H; R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an R 6 configuration. 1 69. The composition of claim 65, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n, m, p and q are 0; R, 3 and R3 are normal fatty acyl residues having 8 carbon atoms; R4, Rs and R7 are H; R 8 4 is phosphono; R, is H; R, and R3 are each attached to a stereogenic center having an R configuration; and R, is attached to a stereogenic center having an S 6 configuration. 1 70. The composition of claim 64, wherein said composition comprises said 2 compound where R6 is H. WO 98/50399 PCT/US98/09385 132 1 71. The composition of claim 70, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n is 2; m, p and q are 0; R,, 3 R 2 and R 3 are normal fatty acyl residues having 14 carbon atoms; R 4 R 5 and R, are H; 4 Rs is phosphono; R 9 is H; and R 2 and R 3 are each attached to a stereogenic center having an R configuration. 1 72. The composition of claim 70, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; n is 1, m and p are 0; q is 3 1; Ri, R 2 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 and R 5 are 4 H; R 7 is carboxy; R 8 is phosphono; R 9 is H; and R, and R 3 are each attached to a stereogenic center having an R configuration. 1 73. The composition of claim 70, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n, p and q are 0; RI, R 2 3 and R 3 are normal fatty acyl residues having 14 carbon atoms; R 4 Rs, and R 7 are H; R 8 4 is phosphono; R 9 is H; and Ri, R 2 and R 3 are each attached to a stereogenic center having an R configuration. 1 74. The composition of claim 70, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n, p and q are 0; R 2 3 and R 3 are normal fatty acyl residues having 10 carbon atoms; R 4 Rs and R 7 are H; R, 4 is phosphono; R9 is H; and R, and R 3 are each attached to a stereogenic center having an R configuration. 1 75. The composition of claim 70, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, p and q are 0; n is 1; R,, 3 R 2 and R 3 are normal fatty acyl residues having 14 carbons; R 4 R 5 and R 7 are H; R 8 is 4 phosphono; R 9 is H; and R, and R 3 are each attached to a stereogenic center having an R configuration. WO 98/50399 PCT/US98/09385 133 1 76. The composition of claim 64, wherein said composition comprises said 2 compound where R 6 is hydroxy. 1 77. The composition of claim 76, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n and q are 0; p is 1; R,, 3 R 2 and R 3 are normal fatty acyl residues having 12 carbon atoms; R 4 and R 5 are H; R 7 4 is H; R 8 is phosphono; and R 9 is H; RI, R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R, is attached to a stereogenic center having an 6 S configuration. 1 78. The composition of claim 76, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m and q are 0; n and p are 3 1; RI, R 2 and R 3 are normal fatty acyl residues having 10 carbon atoms; R4, R 5 and R 7 4 are H; R, is phosphono; R 9 is H; R2 and R 3 are each attached to a stereogenic center having an R configuration; and R, is attached to a stereogenic center having an 6 S configuration. 1 79. The composition of claim 76, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n and q are 0; p is 2; R,, 3 R2 and R3 are normal fatty acyl residues having 10 carbon atoms; R4, R 5 and R7 are H; 4 R8 is phosphono; R 9 is H; R2 and R 3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an S 6 configuration. 1 80. The composition of claim 76, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n and q are 0; p is 1; R,, 3 R 2 and R3 are normal fatty acyl residues having 14 carbon atoms; R 4 R5 and R7 are H; 4 Rg is phosphono; R9 is H; R2 and R3 are each attached to a stereogenic center having an R configuration; and R 5 is attached to a stereogenic center having an R 6 configuration. WO 98/50399 PCT/US98/09385 134 1 81. The composition of claim 76, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n and q are 0; p is 1; R,, 3 R, and R 3 are normal fatty acyl residues having 14 carbon atoms; R, R, and R 7 are H; R, 4 is phosphono; R, is H; R 2 and R, are each attached to a stereogenic center having an R configuration; and R, is attached to a stereogenic center having an S configuration. 1 82. The composition of claim 76, wherein said composition comprises said 2 compound having the following structure: X is 0; Y is O; m, n and q are 0; p is 1; R,, 3 R 2 and R 3 are normal fatty acyl residues having 11 carbon atoms; R, R, and R, are H; Ra 4 is phosphono; R 9 is H; R 2 and R 3 are each attached to a stereogenic center having an 5 R configuration; and R, is attached to a stereogenic center having an S configuration. 1 83. The composition of claim 76, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n and q are 0; p is 1; R,, S. 3 R, and R, are normal fatty acyl residues having 10 carbon atoms; R 4 R5 and R, are H; Rs 4 is phosphono; R, is H; RI, R, and R 3 are each attached to a stereogenic center having an R configuration; and R, is attached to a stereogenic center having an S configuration. 1 84. The composition of claim 76, wherein said composition comprises said 2 compound having the following structure: X is O; Y is O; m, n, p and q are 0; R 2 and 3 R 3 are normal fatty acyl residues having 10 carbon atoms; R, and R, are H; R is amino 4 carbonyl; R7 is H; Rs is phosphono; R, is H; R 2 and R 3 are each attached to a stereogenic center having an R configuration; and R, is attached to a stereogenic center 6 having an S configuration. 1 85. The composition of claim 64, wherein said pharmaceutically acceptable 2 carrier is an aqueous composition comprising water and one or more surfactants selected 3 from the group consisting of glycodeoxycholate, deoxycholate, sphingomyelin, 4 sphingosine, phosphatidylcholine, 1, 2 -Dimyristoyl-sn-glycero-3-phosphoethanolamine, L-a-Phosphatidylethanolamine, and 1, 2 -Dipalmitoyl-sn-glycero-3-phosphocholine, or T a mixture thereof. WO 98/50399 PCT/US98/09385 S135 1
18. 86. The composition of claim 85, wherein said one or more surfactant is 1,2- 2 Dipalmitoyl-sn-glycero-3-phosphocholine. 1
19. 87. The composition of claim 85, wherein the molar ratio of said compound to 2 surfactant is from about 10:1 to 10:5. 1
20. 88. The composition of claim 85, wherein the molar ratio of said compound to 2 surfactant is 4:1. 1
21. 89. The composition of claim 64, wherein said carrier is a stable emulsion r 2 comprising a metabolizable oil, one or more surfactants, an antioxidant and a component 3 to make the emulsion isotonic. 1
22. 90. The composition of claim 89, wherein said stable emulsion comprises S2 v/v squalene, 0.9% w/v PLURONIC-F68 block co-polymer, 1.9% w/v egg phosphatidyl 3 choline, 1.75% v/v glycerol and 0.05% w/v a tocopherol.
23. 91. An inmunoeffector canpound substantially as herein described with reference to the examples.
24. 92. A method for enhancing the inmune response of a manmal substantially as herein described.
25. 93. A vaccine cnposition substantially as herein described.
26. 94. A pharmaceutical canposition substantially as herein described.